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IVo.     17" 

Essentials  of  Diagnosis. 

BY  S.  SOUS-COHEN,  M.  D. 
IVo.    10 

Essentials  of  Hygiene. 

ILLUSTRATED.        BY  ROBERT  P.  ROBIN7S,  M.D.      (In  Preparation.) 

IVo.     SO 

Essentials  of  Bacteriology. 

ILLUSTRATED.  BY  M.  V.  BALL,  M.  D* 

IVo.     2\ 

Essentials  of   Nervous  Diseases  and  Insanity. 

ILLUSTRATED.  BY  JOHN  C.  SHAW,  M.  D. 

IVo.    &2 

Essentials  of  Medical  Physics. 

ILLUSTRATED.       BY  FRED.  J.  BROCKWAY,  M.D.     (In  Preparation.) 

IVo.     23 

essentials  of  Medical  Electricity. 

BY  DAVID  D.  ^TKWART,  M.  D.,  and  EDWARD  S.  LAWRENCE,  M.  D. 
ILLUSTRATED. 

.  leh  CUM  In-  conveniently  curried  in  tin-  pocket,  con  tain 
'  <ichings  of  the  moat  popular  text-boohs. 

;. early  all  connected  with  the  various  colleges  as  Detnon- 

strntnrs',\    r»tiirers,  and  are  therefore  thoroughly  conversant,  not  only  with 
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iiijeets  iii  the  torin  of  questions  and  answers,  will  be  apparent. 

lurd  works,  often   is  :it  a   loss  to  discover 

•  li«-  re  in  em  be  red,  and  is  equally  pn/./,led  w  lien  he  at  tern  pts 

to  the  munuer  in  which  the  questions  could  be  put  in  the 

emaminat  ion-room. 


Ocular 


Nose-piece 


Coarse  Adjustment 
Fine  Adjustment 


Stage 

Abbe  Co 
Iris  Blende 


BACTERIOLOGICAL  MICROSCOPE  (WITH  ABB*  AND  BLENDER  IN  POSITION). 


SAUNDERS'  QUESTION  COMPENDS,  NO.  20. 


ESSENTIALS  OF  BACTERIOLOGY: 


BEING  A 


CONCISE  AND  SYSTEMATIC  INTRODUCTION  TO  THE 
STUDY  OF  MICRO-ORGANISMS 


FOR   THE   USE  OF 


STUDENTS  AND  PRACTITIONERS. 


BY 

M.  V.  BALL,  M.D., 

LATE  RESIDENT  PHYSICIAN  GERMAN  HOSPITAL,   PHILADELPHIA;  ASSISTANT  IN 
MICROSCOPY,  NIAGARA  UNIVERSITY,  BUFFALO,  NEW  YORK,  ETC. 


WITH  SEVENTY-SEVEN  ILLUSTRATIONS,  SOME  IN  COLORS, 


PHILADELPHIA: 

W.    B.    S  AU  N  DE  ES, 

913  WALNUT  STKEET. 
1891. 


COPYRIGHT,  1891. 
BY  W.  B.    SAUNDERS, 


COLLINS  PRINTING  HOUSE, 
705  JAYNE  STREET. 


oo 

PREFACE. 


FEELING  the  need  of  a  Compendium  on  the  subject  of 
this  work,  it  has  been  our  aim  to  produce  a  concise  treatise 
upon  the  Practical  Bacteriology  of  to-day,  chiefly  for  the 
medical  student,  which  he  may  use  in  his  laboratory. 

It  is  the  result  of  experience  gained  in  the  Laboratory 
of  the  Hygienical  Institute,  Berlin,  under  the  guidance  of 
Koch  and  Frankel ;  and  of  information  gathered  from  the 
original  works  of  other  German,  as  well  as  of  French, 
bacteriologists. 

Theory  and  obsolete  methods  have  been  slightly  touched 
upon.  The  scope  of  the  work,  and  want  of  space,  forbade 
adequate  consideration  of  them.  The  exact  measurements 
of  bacteria  have  not  been  given.  The  same  bacterium 
varies  often  much  in  size,  owing  to  differences  in  the  media, 
staining,  etc. 

We  have  received  special  help  from  the  following  books, 
which  we  recommend  to  students  for  further  reference : — 

MAC£:   TraitS  pratique  de  Bacteriologie. 
FRANKEL:    Grundriss  der  Bakterienkunde. 
EISENBERG:    Bakteriologische  Diagnostik. 
CROOKSCHANK,  E.  M.:    Manual  of  Bacteriology. 
GUNTHER:   Einflihriug  in  das  Studium  der  Bacteriologie,  etc. 
WOODHEAD  AND  HARE  :    Pathological  Mycology. 
SALMONSEN:    Bacteriological  Technique  (English  translation). 

M.  V.  BALL. 

BUFFALO,  N.  Y.,  October  1, 1891. 
62  Delaware  Avenue. 

(v) 


CONTENTS. 


PART  I. 
GENERAL  CONSIDERATIONS  AND  TECHNIQUE. 

PAGE 

Introduction •        •        •        .  ix 

CHAPTER  I.— CLASSIFICATION,   STRUCTURE,  AND   RE- 
PRODUCTION .       .       ....  17 

"        II.— ORIGIN,  LIFE,  GROWTH,  AND  PROPERTIES  23 

"       III.— METHODS  OF  EXAMINATION      .       .       .  26 

"       IV.— STAINING  OF  BACTERIA     ....  30 

"         V.— GENERAL  METHOD  OF  STAINING  SPECI- 
MENS        34 

"       VI.— SPECIAL  METHODS  OF  STAINING       .       .37 

"      VII.— METHODS  OF  CULTURE      ....  39 

"    VIII. — NUTRIENT  MEDIA 44 

"       IX.— SOLID  TRANSPARENT  MEDIA     ...  47 

"         X.— INOCULATION  OF  GELATINE  AND  AGAR  .  53 

"       XI.— GROWTH  AND  APPEARANCES  OF  COLONIES  58 

"     XII.— CULTIVATION  OF  ANAEROBIC  BACTERIA  .  60 

"    XIII.— MANNER  IN  WHICH  BACTERIA  ACT  UPON 

BODY 62 

"    XIV.— IMMUNITY 66 

«      XV. — ANIMAL  EXPERIMENTS      ....  68 

(vii) 


viii  CONTENTS. 

PART  II. 
SPECIAL  BACTERIOLOGY. 

PAGE 

CHAPTER  I.— NON-PATHOGENIC  BACTERIA     .       .       .72 

Bacillus  Prodigiosus 72 

Indicus .72 

Mesentericus  Yulgatus 7.'] 

Megatherium 74 

Ramosus 74 

Bacterium  Zopfi 75 

Bacillus  Subtilis 75 

Spinosus 7(3 

Some  Bacteria  in  Milk 76 

Bacillus  Acidi  Lactici 76 

Butyricus 77 

Amylobacter 77 

Lactis  Cyanogenus    ....  78 

Lactis  Erythogenes   ....  78 

Some  Noil-Pathogenic  Bacteria  of  Water    ...  79 

Bacillus  Violaceus ;  Coeruleus         ...  79 

Fluorescent  Bacteria 80 

Phosphorescent  Bacteria         .        .        .        .80 

Crenothrix,  Cladothrix,  and  Beggiatoa    .  81 

Bacterium  Urese 82 

Spirillum 83 

Rubrum ;  Concentricum 83 

Sarcina 83 

Lutea 83 

Aurantica  Flava,  Rosea,  and  Alba  ;  Ventriculi    .  84 

CHAPTER  II.— PATHOGENIC  BACTERIA    .  *     .        .       .84 

Bacteria  Pathogenic  for  Man  and  other  Animals         .  84 

Bacillus  Anthracis 84 

Tuberculosis 88 

Lepra  Bacillus 96 

Syphilis  Bacillus 97 


CONTENTS.  IX 

PAGE 

Bacteria — 

Bacillus  of  Glanders       .        .        .        .        .        .98 

of  Diphtheria 100 

of  Typhoid  Fever' 101 

Neapolitanus 104 

CHAPTER  III.— PATHOGENIC  BACTERIA— CONTINUED    .  105 

Spirillum  Choleras 105 

Bacteria  Similar  to  Spirillum  Cholerse          .        .        .  108 

Finkler-Prior 108 

Tyrogenum 109 

Vibrio  Metschnikoff 110 

Bacteria  of  Pneumonia 110 

Pneumo-bacilltis  of  Friedlander    .        .        .        .111 

of  Frankel  .         .        .        .        .112 

Bacillus  of  Khinoscleroma    „        .        .        .                .  115 

Micrococcus  Tetragenus 115 

Capsule  Bacillus    .        .        .        .        .        .        .  '     .  116 

Micro-Organisms  of  Suppuration          ....  116 

Steptococcus  Pyogenes 117 

Staphylococcus  Pyogenes  Aureus  ....  118 

Micrococcus  Pyogenes  Albus,  Citreus,  Tenuis      .  120 

Cereus  Albus,  Flavus        .        .        .120 

Bacillus  Pyocyaneus 120 

Micrococcus  Gonorrhoea         ....        .  121 

Microbes  Similar  to  Gonorrhoea     ....  123 

Bacillus  of  Tetanus 124 

(Edematis  Maligni 127 

Spirillum  of  Relapsing  Fever         ....  129 

Bacillus  Malarise 130 

Hsematozoa  of  Malaria 130 

CHAPTER  I V.—  BACTERIA  PATHOGENIC  FOR  ANIMALS, 

BUT  NOT  FOR  MAN  .  .  .  .  .      133 

Bacillus  of  Symptomatic  Anthrax        ....    133 

of  Chicken  Cholera          .        .        ......        .134 

Bacteria  of  Hemorrhagic  Septicaemia,  Swine  Plagues, 

Duck  Cholera,  etc 136 


CONTENTS. 

PAGE 

Bacillus  of  Rabbit  Septicaemia     .        .        .        .        .  136 

of  Erysipelas  of  Swine 130 

Murisepticus 138 

Micrococcus  of  Mai  de  Pis 138 

Bacillus  Alvei 139 

Micrococcus  Ainylivorus 139 

Bacterium  Termo  139 


APPENDIX. 

Yeasts 141 

Oidiums 142 

Moulds 143 

Actinomyces  or  Ray  Fungus 144 

Examination  of  Air 146 

of  Water 149 

of  Soil 152 

CONCLUSION  .       .       .  •    .       .  153 


INTRODUCTION. 


HISTORY. — The  microscope  was  invented  about  the  latter 
part  of  the  sixteenth  century ;  and  soon  after,  by  its  aid, 
minute  organisms  were  found  -in  decomposing  substances. 
Kircher,  in  1646,  suggested  that  diseases  might  be  due  to 
similar  organisms ;  but  the  means  at  his  disposal  were  in- 
sufficient to  enable  him  to  prove  his  theories.  Anthony 
Van  Leuwenhoeck,  of  Delft,  Holland  (1680  to  1723),  so 
improved  the  instrument  that  he  was  enabled  thereby  to 
discover  micro-organisms  in  vegetable  infusion,  saliva,  fecal 
matter,  and  scrapings  from  the  teeth.  He  distinguished 
several  varieties,  showed  them  to  have  the  power  of  loco- 
motion, and  compared  them  in  size  with  various  grains  of 
definite  measurement.  It  was  a  great  service  that  this 
"Dutch  naturalist"  rendered  the  world;  and  he  can  rightly 
be  called  the  "  father  of  microscopy." 

Various  theories  were  then  formulated  by  physicians  to 
connect  the  origin  of  different  diseases  with  bacteria ;  but  no 
proofs  of  the  connection  could  be  obtained.  Andry,  in  1701, 
called  bacteria  worms.  Miiller,  of  Copenhagen,  in  1786,  made 
a  classification  composed  of  two  main  divisions — monas  and 
vibrio ;  and  with  the  aid  of  the  compound  microscope  was 
better  able  to  describe  them.  Ehrenberg,  in  1833,  with  still 
better  instruments,  divided  bacteria  into  four  orders:  bac- 
terium, vibrio,  spirillum,  and  spirochsete.  It  was  not  until 
1863  that  any  positive  advance  was  made  in  connecting 
bacteria  with  disease.  Rayer  and  Davaine  had  in  1850 

(xi) 


Xll  INTRODUCTION. 

already  found  a  rod-shaped  bacterium  in  the  blood  of  ani- 
mals suffering  from  splenic  fever  (sang  de  rate),  but  they 
attached  no  special  significance  to  their  discovery  until 
Pasteur  made  public  his  grand  researches  in  regard  to  fer- 
mentation and  the  role  bacteria  played  in  the  economy. 
Then  Davaine  resumed  his  studies,  and  in  1863  established 
by  experiments  the  bacterial  nature  of  splenic  fever  or  an- 
thrax. 

But  the  first  complete  study  of  a  contagious  affection  was 
made  by  Pasteur  in  1869,  in  the  diseases  affecting  silk-worms 
— pebrine  and  flacherie — which  he  showed  to  be  due  to  micro- 
organisms. 

Then  Koch,  in  1875,  described  more  fully  the  anthrax 
bacillus,  gave  a  description  of  its  spores  and  the  properties 
of  the  same,  and  was  enabled  to  cultivate  the  germ  on  arti- 
ficial media ;  and,  to  complete  the  chain  of  evidence,  Pas- 
teur and  his  pupils  supplied  the  last  link  by  reproducing  the 
same  disease  in  animals  by  artificial  inoculation  from  pure 
cultures.  The  study  of  the  bacterial  nature  of  anthrax  has 
been  the  basis  of  our  knowledge  of  all  contagious  maladies, 
and  most  advances  have  been  made  first  with  the  bacterium 
of  that  disease. 

Since  then  bacteriology  has  grown  to  huge  proportions — 
become  a  science  of  itself — and  thousands  of  earnest  workers 
are  adding  yearly  solid  blocks  of  fact  to  the  structure,  which 
structure  it  will  be  our  aim  to  briefly  describe  in  the  pages 
which  are  to  follow, 


ESSENTIALS  OF  BACTERIOLOGY. 

* 

PART  I. 
GENERAL  CONSIDERATIONS. 


CHAPTER  I. 

BACTERIA. 

BACTERIA  (daxtypiov,  little  staff)  is  the  name  given  to  a  group 
of  the  lowest  form  of  plants,  very  closely  following  the  algae. 
They  were  called  Fission- Fungi  or  Schizomycetes  (o^t^w,  to  cleave, 
/uor^j,  fungus),  because  it  was  thought  that,  as  the  Fungi,  they 
lived  without  the  chlorophyll.  The  word  fission  was  supplied 
to  distinguish  them  from  moulds  and  yeasts,  it  denoting  the 
manner  of  reproduction.  Since  several  bacteria  have  been 
found  to  possess  chlorophyll,  and  as  a  great  many  increase  in 
other  ways  than  by  simple  fission— the  name  of  Schizomycetes 
can  no  longer  be  applied,  though  the  word  Bacteria  leaves  much 
to  be  desired. 

Classification.  Ferdinand  Cohn,  in  the  middle  of  the  present 
century,  was  the  first  to  demonstrate  bacteria  to  be  of  vegetable 
origin,  they  being  placed  previous  to  that  among  the  infusoria. 
He  arranged  them  according  to  their  form  under  four  divisions. 

Cohn's  System.    I.  Spherobacteria  (globules). 

II.  Microbacteria  (short  rods).  \    ^ 
III.  Desmobacteria  (long  rods).  / 
IY.  Spirobacteria  (spirals). 
As  expressed  at  the  present  time,  Micrococcus,  Bacillus,  and 
Spirillum.     This  classification  is  very  superficial,  but  because  a 
better  one  has  not  been  found  it  is  most  in  use  to-day. 

2  (17) 


18        ESSENTIALS  OF  BACTERIOLOGY. 

De  Bary's  System.  De  Bary  divides  bacteria  into  two  groups, 
those  arising  from  or  giving  rise  to  endospores  and  those  devel- 
oped from  arthrospores.  This  division  has  a  more  scientific 
value  than  the  first. 

FIG.  1. 


Jo^Tv         ^ 

Micrococcus.  Spirillum.  Bacillus. 

Structure.  Bacteria  are  cells  ;  they  appear  as  round  or  cylin- 
drical of  an  average  diameter  or  transverse  section  of  0.001  mm. 
(=1  micromillimeter),  written  1  /*.  The  cell,  as  other  plant- 
cells,  is  composed  of  a  membranous  cell-wall  and  cell-contents  ; 
"cell-nuclei"  have  not  yet  been  observed,  but  the  latest  re- 
searches point  to  their  presence. 

Cell- Wall.  The  cell-wall  is  composed  of  plant  cellulose,  which 
can  be  demonstrated  in  some  cases  by  the  tests  for  cellulose. 
The  membrane  is  firm  and  can  be  brought  plainly  into  view  by 
the  action  of  iodine  upon  the  cell-contents  which  contracts  them. 

Cell-Contents,  The  contents  of  the  cell  consist  mainly  of 
protoplasm,  usually  homogeneous,  but  in  some  varieties,  finely 
granular,  or  holding  pigment,  chlorophyll,  granulose,  and  sul- 
phur in  its  structure. 

It  is  composed  chiefly  of  myco^yrotcin. 

Gelatinous  Membrane.  The  outer  layer  of  the  cell-membrane 
can  absorb  water  and  become  gelatinoid,  forming  either  a  little 
envelope  or  capsule  around  the  bacterium  or  preventing  the 
separation  of  the  newly-branched  germs,  forming  chains  and 
bunches,  as  strepto-  and  staphylo-cocci.  Long  filaments  are  also 
formed. 

Zoogloea.  "When  this  gelatinous  membrane  is  very  thick,  irre- 
gular masses  of  bacteria  will  be  formed,  the  whole  growth  being 
in  one  jelly-like  lump.  This  is  termed  a  zoogl<wv  (£ww,  animal, 

.  .jlue). 
Locomotion,    Many  bacteria  possess  the  faculty  of  self-move- 


BACTERIA.  19 

ment,  carrying  themselves  in  all  manner  of  ways  across  the 
microscopic  field,  some  very  quickly,  others  leisurely. 

Vibratory  Movements.  Some  bacteria  vibrate  in  themselves, 
appearing  to -move,  but  they  do  riot  change  their  place  ;  these 
movements  are  denoted  as  molecular  or  " Brownian." 

FIG.  2. 


Zooglcea. 

Flagella.  Little  threads  or  lashes  are  found  attached  to  many 
of  the  motile  bacteria,  either  at  the  poles  or  along  the  sides, 
sometimes  only  one,  and  on  some  several,  forming  a  tuft. 

These  flagella  are  in  constant  motion  and  can  probably  be 
considered  as  the  organs  of  locomotion  ;  they  have  not  yet  been 
discovered  upon  all  the  motile  bacteria,  owing  no  doubt  to  our 
imperfect  methods  of  observation.  They  can  be  stained  and 
have  been  photographed.  See  Fig.  3. 

Reproduction.  Bacteria  multiply  either  through  simple  divi- 
sion or  through  fructification  by  means  of  small  round  or  oval 
bodies  called  spores  from  spora-seed.  In  the  first  case,  division, 
the  cell  elongates,  and  at  one  portion,  usually  the  middle,  the 
cell-wall  indents  itself  gradually,  forming  a  septum  and  dividing 
the  cell  into  two  equal  parts,  just  as  occurs  in  the  higher  plant 
and  animal  cells.  See  Fig.  4. 


20 


ESSENTIALS    OF    BACTERIOLOGY. 


FIG.  3. 


Flagella. 

Successive  divisions  take  place,  the  new  members  either  exist- 
ing as  separate  cells  or  forming  part  of  a  community  or  group. 


FJG.  4. 


/-; 


1  Z  3 

Division  of  a  Micrococcus.    (After  Mac6.) 


*._..  — .-* 


/« 


^ 


Division  of  a  Bacillus.    (After  Mace".) 

Spore  Formations.     Two  forms  of  sporulation,  Endosporous 
and  Artlirosporous.    A  small  granule  develops  in  the  protoplasm 


BACTERIA. 


21 


of  a  bacterium,  this  increases  in  size,  or  several  little  granules 
coalesce  to  form  an  elongated,  highly  refractive,  clearly  defined 
object,  rapidly  attaining  its  real  size,  and  this  is  the  spore.  The 
remainder  of  the  cell-contents  has  now  disappeared,  leaving 
the  spore  in  a  dark,  very  resistant,  membrane  or  capsule,  and 
beyond  this  the  weak  cell-wall.  The  cell-wall  dissolves  gradu- 
ally or  stretches  and  allows  the  spore  to  be  set  free. 

Each  bacterium  gives  rise  to  but  one  spore.  It  may  be  at 
either  end  or  in  the  middle  (Fig.  5).  Some  rods  take  on  a  pecu- 
liar shape  at  the  site  of  the  spore,  making  the  rod  look  like  a 
drum-stick  or  spindle,  clostridium  (Fig.  6). 


FIG.  5. 


FIG.  6. 


Sporulation.     After  De  Bary. 


Clostridium. 


Spore  Contents.  What  the  real  contents  of  spores  are  is  not 
known.  In  the  mother  cell  at  the  site  of  the  spore  little  gran- 
ules have  been  found  which  stain  differently  from  the  rest  of 
the  cell,  and  these  are  supposed  to  be  the  beginnings,  the  sporo- 
genic  bodies.  The  most  important  part  of  the  spore  is  its  cap- 
sule; to  this  it  owes  its  resisting  properties.  It  consists  of  two 
separate  layers,  a  thin  membrane  around  the  cell,  and  a  firm 
outer  gelatinous  envelope. 

Germination.  When  brought  into  favorable  conditions,  the 
spore  begins  to  lose  its  shining  appearance,  the  outer  firm  mem- 


22        ESSENTIALS  OF  BACTERIOLOGY. 

brane  begiris  to  swell,  and  it  now  assumes  the  shape  and  size 
of  the  cell  from  which  it  sprang,  the  capsule  having  burst,  so  as 
to  allow  the  young  bacillus  to  be  set  free. 

Requisites  for  Spore  Formation,  It  was  formerly  thought  that 
when  the  substratum  could  no  longer  maintain  it,  or  had  become 
infiltrated  with  detrimental  products,  the  bacterium-cell  pro- 
duced spores,  or  rather  turned  itself  into  a  spore  to  escape  anni- 
hilation ;  but  we  know  now  that  only  when  the  conditions  are 
the  most  favorable  to  the  well-being  of  the  cell,  does  it  produce 
fruit,  just  as  with  every  other  type  of  plant  or  animal  life,  a  cer- 
tain amount  of  oxygen  and  heat  being  necessary  for  good  spore 
formation. 

Asporogenic  Bacteria.  Bacteria  can  be  so  damaged  that  they 
will  remain  sterile,  not  produce  any  spores.  This  condition  can 
be  temporary  only,  or  permanent. 

Arthrosporous.  All  the  above  remarks  relate  to  Endospores, 
spores  that  arise  within  the  cells. 

In  the  other  group  called  Arthrospores,  individual  members 
of  a  colony  or  aggregation  leave  the  same  and  become  the  origi- 
nators of  new  colonies,  thus  assuming  the  character  of  spores. 

The  Micrococci  furnish  examples  of  this  form. 

Some  authorities  have  denied  the  existence  of  the  arthro- 
sporous  formation. 

Resistance  of  Spores.  Because  of  the  very  tenacious  envelope, 
the  spore  is  not  easily  influenced  by  external  measures.  It  is 
said  to  be  the  most  resisting  object  of  the  organic  world. 

Chemical  and  physical  agents  that  easily  destroy  other  life 
have  very  little  effect  upon  it. 

Many  spores  require  a  temperature  of  140°  C.  dry  heat  for 
several  hours  to  destroy  them.  The  spores  of  a  variety  of  potato- 
bacillus  (bacillus  mesentericus)  can  withstand  the  application  of 
steam  at  100°  C.  for  four  hours. 


ORIGIN    OF    BACTERIA.  23 

CHAPTER  II. 

ORIGIN   OF   BACTERIA   AND   THEIR  DISTRIBUTION. 

As  Pasteur  has  shown,  all  bacteria  develop  from  pre-existing 
bacteria,  or  the  spores  of  the  same.  They  cannot,  do  not  arise 
de  novo. 

Their  wide  and  almost  universal  diffusion  is  due  to  the  minute- 
ness of  the  cells  and  the  few  requirements  for  their  existence. 

Very  few  places  are  free  from  germs  ;  the  air  on  the  high  seas, 
and  on  the  mountain  tops,  is  said  to  be  free  from  bacteria, 
but  it  is  questionable. 

One  kind  of  bacterium  will  not  produce  another  kind. 

A  bacillus  does  not  arise  from  a  micrococcus  or  the  typhoid 
fever  bacillus  produce  the  bacillus  of  tetanus. 

This  subject  has  been  long  and  well  discussed,  and  it  would 
take  many  pages  to  state  the  "  pros"  and  "  cons,"  therefore,  this 
positive  statement  is  made,  it  being  the  position  now  held  by  the 
principal  authorities. 

Saprophytes  and  Parasites.  (Saprophytes,  $a?tpo?,  putrid,  $vtov, 
plant.  Parasites,  rtapa,  aside  of  6iro$,  food.)  Those  bacteria 
which  live  on  the  dead  remains  of  organic  life  are  known  as 
Saprophytic  Bacteria,  and  those  which  choose  the  living  bodies 
of  their  fellow-creatures  for  their  habitat  are  called  Parasitic 
Bacteria.  Some,  however,  develop  equally  well  as  Saprophytes 
and  Parasites.  They  are  called  Facultative  Parasites. 

Conditions  of  Life  and  Growth  of  Bacteria.  Influence  of  Tem- 
perature.— In  general,  a  temperature  ranging  from  10°  C.  to  40° 
C.  is  necessary  to  their  life  and  growth. 

Saprophytes  take  the  lower  temperatures  ;  Parasites,  the  tem- 
perature more  approaching  the  animal  heat  of  the  warm-blooded. 
Some  forms  require  a  nearly  constant  heat,  growing  within  very 
small  limits,  as  the  Bacillus  of  Tuberculosis. 

Some  forms  can  be  arrested  in  their  development  by  a  warmer 
or  colder  temperature,  and  then  restored  to  activity  by  a  return 
to  the  natural  heat. 


24:        ESSENTIALS  OF  BACTERIOLOGY. 

A  few  varieties  exist  only  at  freezing  point  of  water ;  and 
others  again  that  will  not  live  under  a  temperature  of  00°  C. 

For  the  majority  of  Bacteria  a  temperature  of  00°  C.  is  de- 
structive ;  and  several  times  freezing  and  thawing  very  fatal. 

Influence  of  Oxygen. — Two  varieties  of  bacteria  in  relation  to 
oxygen. 

The  one  cerofo'c,  growing  in  air  ;  the  other  anccrobic,  living 
without  air. 

Obligate  azrobins,  those  which  exist  only  when  oxygen  is  present. 

Facultative  a>robins,  those  that  live  best  when  oxygen  is  present, 
but  can  live  without  it. 

Obliijate  or  true  anccrobins,  those  which  cannot  exist  where 
oxygen  is. 

facultative  ancerobins,  those  which  exist  better  where  there  is 
no  oxygen,  but  can  live  in  its  presence. 

Some  derive  the  oxygen  which  they  require  out  of  their  nutri- 
ment, so  that  a  bacterium  may  be  terobic  and  yet  not  require 
the  presence  of  free  oxygen. 

JErobins  may  consume  the  free  oxygen  of  a  region  and  thus 
allow  the  anrerobius  to  develop.  By  improved  methods  of  cul- 
ture many  varieties  of  ancerobins  have  been  discovered. 

Influence  of  Light.  —  Sunlight  is  very  destructive  to  bacteria. 
A  few  hours'  exposure  to  the  sun  has  been  fatal  to  anthrax 
bacilli,  and  the  cultures  of  bacillus  tuberculosis  have  been  killed 
by  a  few  days'  standing  in  daylight. 

Effects  of  Electricity. — Electricity  arrests  growth. 

Vital  Actions  of  Microbes.  Bacteria  feeding  upon  organic  com- 
pounds produce  chemical  changes  in  them,  not  only  by  the  with- 
drawal of  certain  elements,  but  also  by  the  excretion  of  these 
elements  changed  by  digestion.  Sometimes  such  changes  are 
destructive  to  themselves,  as  when  lactic  and  butyric  acids  are 
formed  in  the  media. 

Oxidation  and  reducivm  are  carried  on  by  some  bacteria.  Am- 
monia, hydrogen  sulphide,  and  trimethylamin  are  a  few  of  the 
chemical  products  produced  by  bacteria. 

Ptomaines,  Brieger  found  a  number  of  complex  alkaloids, 
closely  resembling  those  found  in  ordinary  plants,  and  which 


ORIGIN    OF    BACTERIA.  25 

he  named  ptomaines,  from  rt^^a  (corpse),  because  obtained 
from  putrefying  objects. 

Fermentation,  This  form  of  "splitting  up"— fermentation, 
as  it  is  called — is  due  to  the  direct  action  of  vegetable  organisms. 
Many  bacteria  have  the  power  of  ferments. 

Putrefaction.  When  fermentation  is  accompanied  by  devel- 
opment of  offensive  gases,  a  decomposition  occurs,  which  is 
called  putrefaction,  and  this,  in  organic  substances,  is  due 
entirely  to  bacteria. 

Liquefaction  of  Solid  Gelatine.  Some  varieties  of  bacteria 
digest  the  nutrient  gelatine,  and  so  dissolve  it;  others  excrete 
a  ferment  which  liquefies  the  gelatine. 

Producers  of  Disease.  Various  pathological  processes  are 
caused  by  bacteria,  the  name  given  to  such  diseases  being 
infectious  diseases  and  the  germs  themselves  called  disease-pro- 
ducing pathogenic  bacteria.  Those  which  do  not  form  any 
pathological  process  are  called  n'.n-pathogenic  bacteria. 

Pigmentation.  Some  of  the  bacteria  are  endowed  with  the 
property  of  forming  pigments  either  in  themselves,  or  producing 
a  chromogenic  body  which,  when  set  free,  gives  rise  to  the  pig- 
ment. In  many  cases  the  pigments  have  been  isolated  and  many 
of  the  properties  of  the  aniline  dyes  discovered  in  them. 

Phosphorescence.  Many  bacteria  have  the  power  to  form 
light,  giving  to  various  objects  which  they  inhabit  a  character- 
istic glow  or  phosphorescence. 

Fluorescence.  An  iridescence  or  play  of  colors  develops  in 
some  of  the  bacterial  cultures. 

Gas  Formation.  Many  bacteria,  anaerobic  ones  especially, 
produce  gases,  noxious  and  odorless  in  the  culture  media,  the 
bubbles  which  arise  soon  displacing  the  media. 

Odors.  Some  germs  form  odors  characteristic  for  them  ;  some 
a  sweet  aromatic  one,  and  others  a  very  foul,  disagreeable  smell ; 
some  give  a  sour  or  rancid  exhalation. 

Effect  of  Age.    With  age,  bacteria  lose  their  strength  and  die. 

Bacteria  thus  carry  on  all  the  functions  of  higher  organized 
life. 

They  breathe,  eat,  digest,  excrete,  and  multiply  ;  and  they  are 
very  busy  workers. 


26        ESSENTIALS  OF  BACTERIOLOGY. 

CHAPTER  III. 

METHODS   OF   EXAMINATION. 

WE  divide  the  further  study  of  the  general  characteristics  of 
Bacteria  into  two  portions  : — 

First  the  examination  of  the  same  by  aid  of  the  microscope. 
Second.  The  continued  study  through  artificial  cultivation. 
They  both  go  hand  in  hand  ;  the  one  incomplete  without  the 
other. 

Microscopical.  The  ordinary  microscope  will  not  suffice  for 
Bacteriological  research.  Certain  special  appliances  must  first 
be  added.  It  is  not  so  much  required  to  have  a  picture  very 
large,  as  to  have  it  sharp  and  clear. 

Oil  Immersion  Lens.  The  penetration  and  clearness  of  a  lens 
are  very  much  influenced  by  the  absorption  of  the  rays  of  light 
emerging  from  the  picture.  In  the  ordinary  dry  system,  many 
of  the  light  rays,  being  bent  outward  by  the  air  which  is  be- 
tween the  object  and  the  lens,  do  not  enter  the  lens,  and  are 
lost.  By  interposing  an  agent  which  has  the  same  refractive 
index  as  glass,  cedar-oil,  or  clove-oil,  for  example,  all  the  rays 
of  light  from  the  object  enter  directly  into  the  lens. 

The  "Homogeneous  System,"  as  this  lens  is  called,  dips  into 
a  drop  of  cedar-oil  placed  upon  the  cover-glass,  and  is  then  ready 
for  use. 

Abbe's  Condenser.    The  second  necessary  adjunct  is  a  com- 
bination  of   mirrors   for    bringing 
FlG-  7'  wide  rays  of  light  directly  under 

the  object.  It  serves  to  intensify 
the  colored  pictures  by  absorbing 
or  hiding  the  unstained  structure. 

This  is  very  useful  in  searching 
a  specimen  for  bacteria,  since  it 
clears  the  field  of  everything  that 
is  not  stained.  It  is  called  Abbe's 

Abbe's  condenser.  Condenser.      Together  with    it  is 

usually  found    an  instrument  for 


METHODS    OF    EXAMINATION.  27 

shutting  off  part  of  the  light— a  Blender.  When  the  bacteria 
have  been  found,  and  their  relation  to  the  structure  is  then 
wished  to  be  studied,  the  "  Abbe"  is  generally  shut  out  by  the 
Iris  blender,  and  the  structure  comes  more  plainly  into  view. 

FIG.  8. 


Iris  Blender. 

For  all  stained  Bacteria  the  oil  immersion  lens  and  Abbe  con- 
denser, without  the  use  of  Blender.  For  unstained  specimens, 
oil  immersion  and  the  narrowed  blender. 

When  examining  with  low  power  objective,  use  a  strong 
ocular.  When  using  high  power  objective  use  weak  ocular.  A 
nose-piece  will  be  found  very  useful,  since  it  is  sometimes  neces- 
sary to  change  the  objective  on  the  same  field,  and  that  insures 
a  great  steadiness  of  the  object. 

Great  cleanliness  is  needed  in  all  bacteriological  methods  ;  but 
nowhere  more  so  than  in  the  microscopical  examination. 

The  cover-glass  should  be  very  carefully  washed  in  alcohol, 
and  dried  with  a  soft  linen  rag.  To  remove  the  stains  on  the 
cover-glasses  that  have  been  used,  they  should  be  soaked  in 
hydrochloric  acid. 

Examination  of  Unstained  Bacteria.    As  the  coloring  of  bac- 


28        ESSENTIALS  OF  BACTERIOLOGY. 

teria  kills  them  and  changes  their  shape  to  some  extent,  it  is  pre- 
ferable to  examine  them  when  possible  in  their  natural  state. 

We  obtain  the  bacteria  for  examination,  either  from  liquid  or 
solid  media. 

From  Liquids.  With  a  long  platinum  needle,  the  end  of  which 
is  bent  into  a  loop,  we  obtain  a  small  drop  from  the  liquid  con- 
taining the  bacteria,  and  place  it  on  a  cover-glass  or  slide  ; 
careful  that  no  bubbles  remain. 

Right  here  we  might  say  that  it  is  best  to  accustom  one's  self 
to  passing  all  instruments,  needles,  etc.,  through  the  flame,  be- 
fore and  after  each  procedure ;  it  insures  safety  ;  and  once  in  the 
habit,  it  will  be  done  automatically. 

From  Solid  Media.  With  a  straight-pointed  platinum  needle, 
a  small  pinch  of  the  medium  is  taken  and  rubbed  upon  a  glass 
slide,  with  a  drop  of  sterilized  water,  or  bouillon,  and  from  this 
a  little  taken  on  cover-glass,  as  before. 

FIG.  9. 


Platinum  Needles. 

The  cover-glass  with  its  drop  is  now  placed  on  the  glass  slide, 
carefully  pressing  out  all  bubbles.  Then  a  drop  of  cedar-oil  is 
laid  on  top  of  the  cover-glass,  and  the  oil  immersion  lens  dipped 
gently  down  into  it  as  close  as  possible  to  the  cover-glass, 
the  narrow  blender  shutting  off  the  Abbe  condenser,  for  this 
being  nn  unstained  specimen,  we  want  but  little  light.  We  now 
apply  the  eye,  and  if  not  in  focus,  use  the  fine  adjustment,  or, 
using  the  coarse,  but  always  away  from  the  object  that  is  to- 
wnrds  us,  since  the  distance  between  the  specimen  and  the  lens 


METHODS    OF    EXAMINATION.  29 

is  very  slight,  it  does  not  require  much  turning  to  break  the 
cover-glass  and  ruin  the  specimen.  Having  found  the  bacte- 
rium, we  see  whether  it  be  bacillus,  micrococcus,  or  spirillum  ; 
discover  if  it  be  motile,  or  not.  That  is  about  all  we  can  ascer- 
tain by  this  method. 

FIG.  10. 


Hanging  Drop  in  Concave  Glass  Slide. 

Hanging  Drop.  When  the  looped  platinum  needle  is  dipped 
into  a  liquid,  a  very  finely-formed  globule  will  hang  to  it ;  this 
can  be  brought  into  a  little  cupped  glass  slide  (an  ordinary 
microscopic  glass  slide  with  a  circular  depression  in  the  centre) 
in  the  following  manner  :  The  drop  is  first  brought  upon  a 
cover-glass ;  the  edges  of  the  concavity  on  the  glass  slide  are 
smeared  with  vaseline,  and  the  slide  inverted  over  the  drop; 
the  cover-glass  sticks  to  the  smeared  slide,  which,  when  turned 
over,  holds  the  drop  in  the  depression  covered  by  the  cover- 
glass,  thus  forming  an  air-tight  cell ;  here  the  drop  cannot 
evaporate. 

Search  for  the  bacteria  with  a  weak  lens  ;  having  found  them, 
place  a  drop  of  cedar-oil  upon  the  cover-glass,  and  bring  the  oil 
immersion  into  place  (here  is  where  a  nose-piece  comes  in  very 
usefully),  careful  not  to  press  against  the  cell,  for  the  cover- 
glasses  are  very  fragile  in  this  position. 

Search  the  edges  of  the  drop  rather  than  the  middle ;  it  will 
usually  be  very  thick  in  the  centre  and  not  so  easily  distin- 
guished. 


30        ESSENTIALS  OF  BACTERIOLOGY. 

Spores,  automatic  movements,  fission,  and  cultivation  in 
general  can  be  studied  for  several  days.  This  moixt  chmnlur 
can  be  placed  in  a  brood-oven  or  on  the  ordinary  warming 
stages  of  the  microscope. 


CHAPTER  IY. 

STAINING   OF   BACTERIA. 

STAINING  or  coloring  bacteria  is  done  in  order  to  make  them 
prominent,  and  to  obtain  permanent  specimens.  It  is  also 
necessary  to  bring  out  the  structure  of  the  bacteria,  and 
serves  in  many  instances  as  a  means  of  diagnosis ;  and  lastly, 
it  would  be  well-nigh  impossible  to  discover'them  in  the  tissues, 
without  staining. 

Only  since  the  aniline  colors  bave  come  into  active  use,  has 
the  technology  of  staining  become  developed. 

Aniline  Colors.  Of  the  numerous  dyes  in  the  market,  nearly 
all  have,  at  one  time  or  other,  been  used  in  staining  bacteria. 
But  now  only  a  very  few  find  general  use,  and  with  methyline 
blue  and  fuchsin  nearly  every  object  can  be  accomplished. 

Basic  and  Acid  Dyes.    Ehrlich  was  the  first  to  divide  the 
aniline  dyes  into  two  groups,  the  basic  colors  to  which  belong— 
Gentian  violet, 
Methyl  violet, 
Methylin  blue, 
Fuchsin, 
Bismark-brown, 
And  the  acid  colors  to  which  eosin  belongs. 

The  basic  dyes  stain  the  bacteria  and  the  nuclei  of  cells  ;  the 
acid  dyes  stain  chiefly  the  tissue,  leaving  the  bacteria  almost 
untouched.  Carmine  and  Hcematoxylin  are  also  useful  as  con- 
trast stains,  affecting  bacteria  very  slightly.  The  aniline  dyes 
are  soluble  in  alcohol  or  water  or  a  mixture  of  the  two. 

Staining  Solutions.  A  saturated  solution  of  the  dye  is  made 
with  alcohol.  This  is  called  the  stock  or  concentrated  solution ; 


STAINING    OF    BACTERIA.  31 

1  part  of  this  solution  to  about  100  parts  of  distilled  water  con- 
stitutes the  ordinary  aqueous  solution  in  use  or  weak  solution. 

It  is  readily  made  by  adding  to  an  ounce  bottle  of  distilled 
water  enough  of  the  strong  solution  until  the  fluid  is  just 
opaque  in  the  body  of  the  bottle,  but  still  clear  in  the  neck  of 
the  same. 

These  weak  solutions  should  be  renewed  every  three  or  four 
weeks,  otherwise  the  precipitates  formed  will  interfere  with  the 
staining. 

Compound  Solutions.  By  means  of  certain  chemical  agents, 
the  intensity  of  the  aniline  dyes  can  be  greatly  increased. 

Mordants.  Agents  that  "frrte"  into  the  specimen  carrying 
the  stain  with  them,  depositing  it  in  the  deeper  layers,  are 
called  mordants  or  etchers. 

Various  metallic  salts  and  vegetable  acids  are  used  for  such 
purpose. 

The  mother  liquid  of  the  aniline  dyes,  aniline  oi7,  a  member 
of  the  aromatic  benzol  group,  has  also  this  property. 

Aniline  Oil  Water.  Aniline  oil  is  shaken  up  with  water  and 
then  filtered  ;  the  aniline  water  so  obtained  is  mixed  with  the 
dyes  forming  the  "aniline  water  gentian  violet"  or  aniline 
water  fuchsin,  etc. 

Carbol  Fuchsin.  Carbolic  acid  can  be  used  instead  of  aniline 
oil,  and  forms  one  of  the  main  ingredients  of  Ziehl's  or  Neelsen's 
solution,  used  principally  in  staining  bacillus  tuberculosis. 
Kiihne  has  a  carbol-methylin  blue  made  similar  to  the  carbol 
fuchsin. 

Alkaline  Stains.  Alkalies  have  the  same  object  as  the  above 
agents ;  namely,  to  intensify  the  picture.  Potassium  hydrate, 
ammon.  carbonate,  and  sodium  hydrate  are  used. 

Loffler's  alkaline  blue  and  Koch's  weak  alkaline  blue  make 
use  of  potassium. 

Heat.  Warming  or  boiling  the  stains  during  the  process  of 
staining  increases  their  intensity. 

Decolorizing  Agent's.  The  object  is  usually  over-colored  in 
some  part,  and  then  decolorizing  agents  are  employed.  Water  is 
sufficient  for  many  cases  ;  alcohol  and  strong  mineral  acids  com- 
bined are  necessary  in  some. 


32        ESSENTIALS  OF  BACTERIOLOGY. 

Iodine  as  used  in  Gram's  Method.  Belonging  to  this  group, 
but  used  more  in  the  sense  of  a  protective,  is  tr.  iodine.  It  picks 
out  certain  bacteria,  which  it  coats  ;  prevents  tliem  from  being 
decolorized,  but  allows  all  else  to  be  faded.  Then  by  using  one 
of  the  acid  or  tissue  dyes,  a  contrast  color,  or  double  staining  is 
obtained.  Many  of  the  more  important  bacteria  are  not  acted 
upon  by  the  iodine,  and  it  thus  becomes  a  very  useful  means  of 
diagnosis. 

Formulas  of  different  Staining  Solutions. 

I. — Saturated  Alcoholic  Solution. 

Place  about  10  grammes  of  the  powdered  dye  in  a  bottle  and 
add  40  grammes  of  alcohol.  Shake  well  and  allow  to  settle. 
This  can  be  used  as  the  stock  bottle. 

II.—  Weak  Solutions. 

Made  best  by  adding  about  1  part  of  number  I.  or  stock  solu- 
tion to  10  of  distilled  water.  This  is  the  ordinary  solution  in  use. 

III.— Aniline  Oil  Water. 

Aniline  oil 5  parts. 

Distilled  water        ....     100  parts.— M. 
Shake  well  and  filter.     To  be  made  fresh  each  time. 

IV. — Aniline  Water  Dyes. 

Sat.  alcoh.  sol.  of  the  dye       .        .  11  parts. 

Aniline  oil  water    ....  100  parts. 

Abs.  alcohol 10  parts. — M. 

Can  be  kept  10  days. 

V.—  Alkaline  Methylin  Jllue. 

A.  J30fer*«. 

Sat.  ale.  sol.  methylin  blue     .         .       30 
Sol.  potass,  hydrat.  (1-10,000)         .     100— M. 

B.  KodSs. 

Sol.  potass,  hydrat.  (10  per  cent.)  0.2 

Sat.  ale.  sol.  methyl,  blue       .         .         1.0 
Distilled  water  200.0— M. 


STAINING    OF    BACTERIA.  33 

VI. —  Carbolic  Acid  Solutions. 

A.  Ziehl-Neelsen. 

Fuchsia  (powd.)      ....  1  part. 

Alcohol 10  parts. 

5  per  cent.  sol.  acid,  carbolic          .  100  parts.— M. 
Filter.     The  older  the  solution  the  better. 

B.  Kuhne. 

Methylin  blue          ....        1.5 

Alcohol 10.0 

5  per  cent.  sol.  ac.  carbol.       .         .     100.0 
Add  the  acid  gradually.     Tins  solution  loses  strength  with  age. 

VII. — Gram's  Iodine  Solution. 
Iodine     ......        1 

Potass,  iod 2 

Aquae  destillat.        .         .         .         .     300.— M. 

VIII.— Lqffler's  Mordant  (for  flagella). 
Aq.  sol.  of  tannin  (20  per  cent.)     .       10  parts. 
Aq.  sol.  ferri  sulph.  (5  per  cent.)    .        1  part. 
Aqure  decoc.  of  logwood  (1-8)        .        4  parts. — M. 
Keep  in  well-corked  bottle. 

IX. — Picro-carmine  ( Ranvier). 

Carmine 1 

Water 10 

Sol.  atnmon.    .....        3 

Sat.  sol.  picric  acid          .        .        .    200.— M. 

X. — Gobbet's  Acid  Blue  (rapid  stain). 
Methylin  blue          ....        2 
25  per  cent,  sulphuric  acid      .        .     100. — M. 

XI. — Alkaline  Aniline  Water  Solutions. 
Sodium  hydrat.  (1  per  cent.)  .        .        1 
Aniline  oil  water    ....     100. — M. 
And  add — 

Fuchsin,  or  methyl-violet  powd.    .        4 
Cork  well.     Filter  before  using. 

O 


34        ESSENTIALS  OF  BACTERIOLOGY. 

CHAPTER  V. 

GENERAL   METHOD   OF   STAINING   SPECIMENS. 

Cover-Glass  Preparations.  The  material  is  evenly  spread  in 
as  thin  a  layer  as  possible  upon  a  cover-glass ;  then,  to  spread 
it  still  more  finely,  a  second  cover-glass  is  pressed  down  upon 
the  first  and  the  two  slid  apart.  This  also  secures  two  speci- 
mens. Before  they  can  be  stained  they  must  be  perfectly  dry, 
otherwise  deformities  will  arise  in  the  structure. 

Drying  the  Specimen.— The  cover-glass  can  be  set  aside  to  dry 
or  held  in  the  fingers  over  the  Bunsen  burner  (the  fingers  prevent- 
ing too  great  a  degree  of  heat).  Since  most  of  the  specimens 
contain  a  certain  amount  of  albumenoid  material,  it  is  best  in 
all  cases  to  "  fix"  it,  i.  e.,  to  coagulate  the  albumen.  This  is 
accomplished  by  passing  the  cover-glass  (after  the  specimen  is 
dry)  three  times  through  the  flame  of  the  burner,  about  three 
seconds  being  consumed  in  doing  so,  the  glass  being  held  in  a 
small  forceps,  smeared  side  up. 

The  best  forceps  for  grasping  cover-glasses  is  a  bent  one,  bent 
again  upward,  near  the  ends.  •* 

The  object  is  now  ready  for  staining. 

Staining. — A  few  drops  of  the  staining  solution  are  placed 
upon  the  cover-glass  so  that  the  whole  specimen  is  covered, 
and  it  is  left  on  a  few  minutes,  the  time  depending  upon  the 
variety,  the  strength  of  stain,  and  the  object  desired.  Instead 
of  placing  the  dye  upon  the  object,  the  cover-glass  can  be  im- 
mersed in  a  small  glass  dish  containing  the  solution  ;  or,  if 
heat  is  desired  to  intensify  or  hasten  the  process,  a  watch- 
crystal  holding  the  stain  is  placed  over  a  Bunsen  burner  and 
in  it  the  cover-glass ;  and,  again,  the  cover-glass  can  be  held 
directly  in  the  flame  with  the  staining  fluid  upon  it,  which 
must  be  constantly  renewed  until  the  process  is  completed. 

Removing  Excess  of  Stain.  The  surplus  stain  is  washed  oil* 
by  dipping  the  glass  in  water,  distilled  water  always  best, 
though  ordinary  running  water  is  admissible. 

The   water  is  removed    by  drying    between   filter  paper  or 


METHOD  OF  STAINING  SPECIMENS.       35 

simply  allowed  to,  run  off  by  standing  the  cover-glass  slant- 
wise against  an  object.  When  the  specimen  is  to  be  examined 
in  water  (which  is  always  best  with  the  first  preparation  of 
the  specimen,  as  the  Canada  balsam  destroys  to  some  extent 
the  natural  appearance  of  the  bacteria),  a  small  drop  of  ster- 
ilized water  is  placed  upon  the  glass  slide,  and  the  cover-glass 
dropped  gently  down  upon  it,  so  that  the  cover-glass  remains 
adherent  to  the  slide. 

The  dry  system  or  the  oil-immersion  can  now  be  used. 

When  the  object  has  been  sufficiently  examined  it  can  be  per- 
manently mounted  by  lifting  the  cover-glass  off  the  slide  (this 
is  facilitated  by  letting  a  little  water  flow  under  it,  one  end 
being  slightly  elevated).  The  water  that  still  adheres  is  dried 
off  in  the  air  or  gently  over  the  flame,  and  when  perfectly  dry 
it  is  placed  upon  the  drop  of  Canada  balsam  which  has  been  put 
upon  the  glass  slide. 

In  placing  the  cover-glass  in  the  staining  solutions  one  must 
be  careful  to  remember  which  is  the  spread  side. 

By  holding  it  between  yourself  and  the  window,  and  scraping 
the  sides  carefully  with  the  sharp  point  of  the  forceps,  the  side 
having  the  specimen  on  it  will  show  the  marks  of  the  instrument. 

Little  glass  dishes,  about  one-half-dozen,  should  be  at  hand 
for  containing  the  various  stains  and  decolorants.. 

Tissue  Preparations.  In  order  to  obtain  suitable  specimens 
for  staining,  very  thin  sections  of  the  tissue  must  be  made. 

As  with  histological  preparations,  the  tissue  must  be  hardened 
before  it  can  be  cut  thin  enough.  Alcohol  is  the  best  agent  for 
this  purpose. 

Pieces  of  the  tissue  one-quarter  inch  in  size  are  covered  with 
alcohol  for  24  to  48  hours. 

When  hardened  it  must  be  fixed  upon  or  in  some  firm  object. 
A  paste  composed  of — 

Gelatine       ......     1  part. 

Glycerine 4  parts. 

Water 2  parts. 

will  make  it  adhere  firmly  to  a  cork  in  about  2  hours,  or  it  can 
be  imbedded  in  a  small  block  of  paraffine,  and  covered  over  with 
melted  paraffine. 


3t>  ESSENTIALS    OF    BACTERIOLOGY. 

Cutting.     The  microtome  should  be  able  to  cut  sections 
inch  in  thickness  ;  this  is  the  fineness  usually  required. 

The  sections  are  brought  into  alcohol  as  soon  as  cut  unless 
they  have  been  imbedded  in  paraffine,  when  they  are  first  washed 
in  chloroform  to  dissolve  out  the  paraffine. 

Staining.  All  the  various  solutions  should  be  in  readiness, 
best  placed  in  the  little  dishes  in  the  order  in  which  they  are  to 
be  used,  as  a  short  delay  in  one  of  the  steps  may  spoil  the  speci- 
men. 

A  very  useful  instrument  for  transferring  the  delicate  sections 
from  one  solution  to  another  is  a  little  metal  spatula,  the  blade 
being  flexible. 

A  still  better  plan,  especially  when  the  tissue  is  "crumbling," 
is  to  "carry  out"  the  whole  procedure  on  the  glass-side. 

General  Principles.  The  section  is  transferred  from  the  alco- 
hol in  which  it  has  been  kept  into  water,  which  removes  the 
excess  of  alcohol,  from  here  into — 

Dish  I,  containing  the  stain;  where  it  remains  5  to  15  minutes. 
Then- 

Dith  II,  containing  5  per  cent,  acetic  acid  (1  to  20) ;  where  it 
remains  £  to  1  min.  The  acid  removes  the  excess  of  stain. 

Dish  III,  water  to  rinse  off  the  acid.  The  section  can  now  be 
placed  under  the  microscope  covered  with  cover-glass  to  see  if 
the  intensity  of  the  stain  is  sufficient  or  too  great.  A  second 
section  is  then  taken,  avoiding  the  errors,  if  any ;  and  having 
reached  this  stage  proceeded  Avith  as  follows  : — 

Dish  IV,  alcohol,  2  to  3  seconds  to  remove  the  water  in  the 
tissue. 

V.  A  few  drops  of  oil  of  clores,  just  long  enough  to  clear  the 
specimen  to  make  it  transparent  (so  that  an  object  placed  under- 
neath will  shine  through). 

VI.  Remove  excess  with  filter-paper. 

VII.  Mount  in  Canada  balsam. 


STAINING    AND    MODIFICATIONS.  37 

CHAPTER  VI. 

SPECIAL  METHODS  OF  STAINING  AND  MODIFICATIONS. 

Gram's  Method  of  Double  Staining.  (For  cover-glass  speci- 
mens.)— I.  A  hot  solution  of  anil,  water  gentian  violet  2  to  10 
minutes. 

II.  Directly  without  washing,  into  Gram's  solution  of  iod. 
potass,  iod.  1  to  3  min.  (the  cover-glass  looks  black). 

III.  Wash  in  alcohol  60  per  cent,  until  only  a  light  brown 
shade  remains  (as  if  the  glass  were  smeared  with  dried  blood). 

IV.  Ilinse  off  alcohol  with  water. 

V.  Contrast  color  with  either  eosin,  picro-carmine,  orbismark- 
brown.     The  bacteria  will  appear  deep  blue,  all  else  red  or  brown 
on  a  very  faint  brown  background. 

The  following  bacteria  do  not  retain  their  color  with  Gram's 
method — are  therefore  not  available  for  the  stain  : — 
Bacillus  of  typhoid. 
Spirillum  of  cholera. 
Bacillus  of  chicken  cholera. 
"        of  rabbit  septica3mia. 
"        of  malignant  O3dema. 
"        of  pneumonia  (Friedlander). 
"        of  glanders. 
Diplococcus  of  gonorrhoea. 
Spirillum  of  relapsing  fever. 
Gram's  Method  for  Tissues  (modified  by  GUnther). 

I.  Stain  in  anil,  water  gent,  violet      .        .     1  minute. 

II.  Dry  between  filter  paper. 

III.  Iod.  potass,  iod.  sol.         .         .         .         .2  minutes. 

IV.  Alcohol ^  minute. 

V.  3  perct.  sol.  hydrochloric  acid  in  alcohol    10  seconds. 

VI.  Alcohol,  ol.  of  cloves,  and  Canada  balsam. 

To  Stain  Spores.  Since  spores  have  a  very  firm  capsule, 
which  tends  to  keep  out  all  external  agents,  a  very  intensive 
stain  is  required  to  penetrate  them,  but  once  this  object  attained 
it  is  equally  as  difficult  to  decolorize  them. 


38  ESSENTIALS    OF    BACTERIOLOGY. 

A  cover-glass  prepared  in  the  usual  way,  i.  e.,  drying  and 
passing  the  specimen  through  the  flame  three  times,  is  placed  in 
a  watch-crystal  containing  Ziehl's  carbol-t'uchsin  solution,  and 
the  same  placed  upon  a  rack  over  a  Bunsen  burner,  where  it  is 
kept  at  boiling-point  for  one  hour,  careful  to  supply  fresh  solution 
at  short  intervals  lest  it  dry  up. 

The  bacilli  are  now  decolorized  in  alcohol,  containing  £  per 
cent,  hydrochloric  acid.  A  contrast  color,  preferably  methyliu 
blue,  is  added  for  a  few  minutes. 

The  spores  will  appear  as  little  red  beads  in  the  blue  bacteria, 
and  loose  ones  lying  about. 

Spore  Stain  (modified). — I.  Carlol.-fuchsin  on  cover-glass  and 
heated  in  the  flame  to  boiling  point  20  to  30  times. 

II.  25  per  cent,  sulphuric  acid,  2  seconds  ;  rinsed  in  water. 

III.  Methylin  blue  contrast. 

Flagella  Stain,  with  Loffkr^s  Mordant.— I.  A  few  drops  of  the 
mordant  (No.  viii.)  are  placed  upon  the  spread  cover-glass  and 
heated  until  it  steams. 

II.  Washed  with  water  until  the  cover-glass  looks  almost  clean, 
using  a  small  piece  of  filter  paper  to  rub  off  the  crusts  which  have 
gathered  around  the  edges. 

III.  Aniline  water  fuchsin  (neutral)  held  in  flame  about  H 
minutes. 

IV.  Wash  in  water. 

If  the  stain  is  properly  made,  the  microbes  are  deeply  colored 
and  the  flagella  seen  as  little  dark  lines  attached  to  them. 

Spoi'ogenic  bodies  stain  quite  readily,  and  in  order  to  distin- 
guish them  from  spores  Ernst  uses  alkaline  nuthylin  blue,  slightly 
warmed. 

Then  rinse  in  water. 

Contrast  with  cold  bismark -brown. 

The  spores  are  colored  bright  blue,  the  spore  granules  a  dirty 
blue,  being  mixed  with  the  brown,  which  colors  also  the  bacteria. 

Kiihntfs  Method. — In  sections,  the  alcohol  used  sometimes  de- 
colorizes too  much.  To  obviate  this  K-tihm  mixes  the  alcohol 
with  the  stain,  so  that  while  the  section  is  beting  anhydrated  it 
is  constantly  supplied  with  fresh  dye. 

Weiyert  uses  aniline  oil  to  dehydrate  instead  of  alcohol,  and 
here,  too,  it  can  be  used  mixed  with  the  dye. 


METHODS    OF    CULTURE.  39 

General  Double  Staining  for  Sections. 
I.  Stain  (watery  dyes)        .         .        .  10  to  15  minutes. 
II.  Acetic  acid  and  water  (1  to  4)       .       £  minute. 

III.  Alcohol 2  to  3  minutes. 

IV.  Contrast  stain,  usually  picro-car- 

mine  or  eosiu      .        .         .        .     2  to  3  minutes. 

Y.  Alcohol i  minute. 

VI.  Clove  oil.     Canada  balsam. 

Instead  of  coloring  with  the  contrast  last,  it  can  be  used  first, 
then  alcohol  one-half  minute,  followed  by  the  bacteria  stain,  acid 
water,  alcohol,  clove  oil,  and  Canada  balsam  in  succession. 

The  stains  for  special  bacteria  will  be  given  when  treating  of 
the  same. 


CHAPTER  VII. 

METHODS   OF   CULTURE. 

Artificial  Cultivation.— The  objects  of  cultivation  are  to  obtain 
germs  in  pure  culture,  free  from  all  foreign  matter,  isolated  and 
so  developed  as  to  be  readily  used  either  for  microscopical  ex- 
amination or  animal  experimentation. 

To  properly  develop  bacteria  we  supply  as  near  as  possible 
the  conditions  which  hold  for  the  especial  germ  in  nature. 
With  the  aid  of  solid  nutrient  media  the  bacteria  can  be  easily 
separated,  and  the  methods  are  nearly  perfect. 

Sterilization,  If  we  place  our  nutrient  material  in  vessels 
that  have  not  been  properly  disinfected,  we  will  obtain  growths 
of  bacteria  without  having  sown  any. 

If  we  have  thoroughly  cleaned  our  utensils,  and  then  not  taken 
care  to  protect  them  from  further  exposure,  the  germs  we  have 
sown  will  be  effaced  or  contaminated  by  multitudes  of  others, 
that  are  constantly  about  us.  We  therefore  have  two  neces- 
sary precautions  to  take  : — 

First.  To  thoroughly  clean  and  sterilize  every  object  that 
enters  into,  or  in  any  way  comes  in  contact  with,  the  culture. 


40 


ESSENTIALS  OF  BACTERIOLOGY. 


Second.  To  maintain  this  degree  of  disinfection  throughout 
the  whole  course  of  the  growth,  and  prevent,  by  proper  con- 
tainers, the  entrance  of  foreign  germs. 

Disinfectants.  Corrosive  sublimate  (bichloride  of  mercury), 
which  is  the  most  effective  agent  we  possess,  cannot  be  gene- 
rally used  because  it  renders  the  soil  unproductive  and  therefore 
must  only  be  employed  in  washing  dishes,  to  destroy  the  old 
cultures.  Even  after  washing,  a  few  drops  of  the  solution  may 
remain  and  prevent  growth,  so  that  one  must  be  careful  to  have 
the  glass-ware  that  comes  in  contact  with  the  nutrient  media 
not  too  moist  with  the  sublimate. 

Heat.  Heat  is  the  best  agent  we  possess  for  general  use. 
Dry  heat  and  moist  heat  are  the  two  forms  employed. 

FIG.  11. 


Hot  Air  Oven. 

For  obtaining  dry  heat— that  is,  a  temperature  of  150°  C., 
(about  300°  F.) — a  sheet-iron  oven  is  used  which  can  be  heated 
by  a  gas-burner.  If  it  have  double  walls  (air  circulating  be- 
tween), the  desired  temperature  is  much  more  quickly  obtained. 
A  small  opening  in  the  top  to  admit  a  thermometer  is  neces- 
sary. These  chests  are  usually  about  1  foot  high,  1|  foot  wide, 


METHODS    OF    CULTURE. 


41 


and  |  foot  deep.  In  them,  glass-ware,  cotton,  and  paper  can  be 
sterilized.  When  the  cotton  is  turned  slightly  brown  it  usually 
denotes  sufficient  sterilization.  All  instruments,  where  prac- 
ticable, should  be  drawn  through  flame  of  alcohol  lamp  or  Bun- 
sen  burner. 

Moist  Heat. — Steam  of  100°  C.  in  circulation  has  been  shown 
to  be  a  very  effective  application  of  heat. 


Koch's  Steam-chest,  The  best 
way  of  obtaining  circulating 
steam  is  by  aid  of  Koch's  appa- 
ratus. This  consists  of  a  cylin- 
drical tin  chest  about  2£  feet 
high  and  about  |  foot  in  diame- 
ter ;  divided  in  its  interior  by  a 
perforated  diaphragm,  a,  an  up- 
per chamber  for  the,  c,  steam  and 
a  lower  one  for  water,  6.  Two 
or  more  gas-burners  placed  under- 
neath the  chest,  which  stands  on 
a  tripod,  supply  the  heat.  In  the 
cover  is  an  opening  for  a  ther- 
mometer. The  chest  is  usually 
covered  with  felt.  When  the 
thermometer  registers  100°  C,, 
the  culture  medium  or  other  sub- 
stance to  be  sterilized  is  placed 
in  the  steam  and  kept  there  10  to 
15  minutes,  or  longer,  as  required. 

The  autoclave  of  Chamberland 
allows  a  temperature  of  120°  C, 
to  be  obtained,  and  is  much  used 
in  Pasteur's  laboratory. 

Instead  of  sterilizing  for  a  long 
time  at  once,  successive  steriliza- 
tion is  practised  with  nutrient 
media,  so  that  the  albumen  will 
not  be  too  strongly  coagulated, 
three  days  in  succession. 


FIG.  12. 


Koch's  Steam-chest. 


Fifteen  minutes  each  day  for 


42        ESSENTIALS  OF  BACTERIOLOGY. 

Fin.  13. 


Chamberland's  Autoclave  with  pressure. 

Fractional  Sterilization  of  Tyndall.  Granted  that  so  many 
spores  originally  exist  in  the  object  to  be  sterilized,  it  is  sub- 
jected to  60°  C.  for  four  hours,  in  which  time  a  part  at  least  of 
those  spores  have  developed  into  bacteria,  and  the  bacteria 
destroyed  by  the  further  application  of  the  heat.  The  next  day 
more  bacteria  will  have  formed,  and  four  hours'  subjection  to 
GO0  heat  will  destroy  them,  and  so  at  the  end  of  a  week,  using 


METHODS    OF    CULTURE. 


43 


four  hours'  application  each  day,  all  the  spores  originally  present 
will  have  germinated  aud  the  bacteria  destroyed. 

Cotton  Plugs  or  Corks.  All  the  glass  vessels  (test-tubes,  flasks, 
etc.)  must  be  closed  with  cotton  plugs,  the  cotton  being  easily 
sterilized  and  preventing  the  entrance  of  germs. 

Test-tubes.  New  test-tubes  are  washed  with  hydrochloric 
acid  and  water  to  neutralize  the  alkalinity  often  present  in 
fresh  glass.  They  are  then  well  washed  and  rubbed 'with  a 
brush,  placed  obliquely  to  drain,  and  when  dry  corked  with 


FIG.  14. 


FIG.  15. 


Wire-Cage. 


Cotton  plugged  Test-Tubes. 


cotton  plugs.  Then  put  in  the  hot-air  oven  (little  wire-cages 
being  used  to  contain  them)  for  fifteen  minutes,  after  which  they 
are  ready  to  be  filled  with  the  nutrient  media.  (The  cotton 
should  fit  firmly  in  the  tube  and  extend  a  short  space  beyond  it. ) 
Test-tubes  without  flaring  edges  are  more  desirable  since  the 
edges  can  easily  be  drawn  out  so  as  to  seal  the  tube. 


44  ESSENTIALS    OF    B  A  CTERIOLOG  Y  . 

CHAPTER  VIII. 

NUTRIENT   MEDIA. 

OF  the  many  different  media  recommended  and  used  since 

bacteriology  becaiwe  a  science,  we  can  only  describe  the  more 

important  ones  now  in  use.     Each  investigator  changes  them 
according  to  his  taste. 

FLUID  MEDIA. 

Bouillon  (according  to  Loffler).  A  cooked  infusion  of  chopped 
beef  made  slightly  alkaline  with  carbonate  of  soda.  Prepared 
as  follows  :  500  grammes  of  finely-chopped  raw  lean  beef  are 
placed  in  a  wide-mouthed  jar  and  covered  with  1  litre  of  water  ; 
this  is  left  standing  twelve  hours  with  occasional  shaking.  It  is 
then  strained  through  cheese  cloth  or  straining  cloth,  the  white 
meat  remaining  in  the  cloth  being  pressed  until  one  litre  of  the 
blood  red  meat-water  has  been  obtained.  The  meat-water  must 
now  be  cooked,  but  before  doing  this,  in  order  to  prevent  all  the 
albumen  from  coagulating,  10  parts  of  peptone  powder  and  5 
parts  of  common  salt  are  added  to  every  1000  parts  meat-water. 
It  is  next  placed  in  the  steam-chest  or  water-bath  for  three- 
quarters  of  an  hour. 

Neutralization.  The  majority  of  bacteria  grow  best  on  a 
neutral  or  slightly  alkaline  soil,  and  the  bouillon,  as  well  as 
other  media,  must  be  carefully  neutralized  with  a  sat.  sol.  of 
carbonate  of  soda.  Since  too  much  alkalinity  is  nearly  as  bad 
as  none  at  all.  the  soda  must  be  added  drop  by  drop  until  red 
litmus  paper  commences  to  turn  blue.  The  bouillon  is  then 
cooked  another  hour,  and  liltered  when  cold.  The  liquid  thus 
obtained  must  be  clearly  alkaline,  and  not  clouded  by  further 
cooking.  If  cloudiness  occur,  the  white  of  an  egg  and  further 
boiling  will  clear  the  same. 

Sterilization  of  the  bouillon.  Erlenmeyer  flasks  (little  conical 
glass  bottles)  or  test-tubes  plugged  and  properly  sterilized  are 
filled  one-third  full  with  the  bouillon,  and  placed  with  their  con- 
tents in  the  steam-chest.  A  tin  pail  with  perforated  bottom 


NUTRIENT    MEDIA.  <J5 

makes  a  good  container  in  which  they  can  be  lowered  in  the 
Koch's  oven.  They  are  left  in  steam  of  100°  C.  one  hour  for 
three  successive  days,  after  which  the  tubes  and  bouillon  are 
ready  for  use. 

Solid  Media.  The  knowledge  of  bacteria  and  germs  of  moulds 
settling  and  growing  upon  slices  of  potato  exposed  to  the  air,  led 
to  the  use  of  solid  media  for  the  artificial  culture  of  the  same, 
It  was  also  thus  learned  that  each  germ  tends  to  form  a  separate 
colony  and  remain  isolated, 

Potato-Cultures.  A  ripe  potato  with  a  smooth  skin  is  the 
best. 

Several  are  brushed  and  scrubbed  with  water  to  get  rid  of  the 
dirt  and  the  *'eyes"  are  cut  out. 

Next  placed  in  1  to  500  solution  of  bichloride  of  mercury  for 
|  hour,  Then  in  the  steam-chest  for  f  hour. 

In  the  meantime,  a  receptable  is  prepared  for  them.  This  is 
called  the  moist  chamber. 

The  moist  chamber  consists  of  two  large  shallow  dishes,  one, 
the  larger,  as  a  cover  to  the  other. 

These  dishes  are  washed  in  warm  distilled  water. 

A  layer  of  filter  paper  moistened  with  a  15  to  30  drops  of  1  to 
1000  bichloride  is  placed  in  the  bottom  of  the  glass  dish. 

FIG.  16. 


Moist  chamber  for  potatoes. 

The  operator  now  prepares  his  own  hands,  rolling  up  his  coat 
sleeves  and  carefully  washing  his  hands,  then  taking  a  potato 
from  the  steam-oven  and  holding  it  between  his  thumb  and 
index  finger  in  the  short  axis,  he  divides  the  potato  in  its 
long  axis  with  a  knife  that  has  been  passed  through  the  flame. 
The  two  halves  are  kept  in  contact  until  they  are  lowered  into 


46        ESSENTIALS  OF  BACTERIOLOGY. 

the  moist  chamber,  when  they  of  their  own  weight  fall  aside, 
the  cut  surface  uppermost.     They  are  then  ready  for  inoculation. 

Fio.  17. 


Method  of  slicing  potato.    (After  Woodhead  and  Hare.) 

Esmarch's  Cubes.  The  potato  is  first  well  cleaned  and  peeled. 
It  is  then  cut  in  cubes  £  inch  in  size. 

These  are  placed,  each  in  a  little  glass  dish  or  tray  and  then 
in  steam-chest  for  £  hour,  after  which  they  are  ready  for  inocu- 
lation (the  dishes  first  having  been  sterilized  in  hot-air  oven). 

Test-tube  Potatoes.  Cones  are  cut  out  of  the  peeled  potato 
and  placed  in  test-tubes,  which  can  then  be  plugged  and  easily 
preserved. 

Manner  of  Inoculation.  With  a  platinum  rod  or  a  spatula 
(sterilized)  the  material  is  spread  out  upon  one  of  the  slices, 
keeping  free  of  the  edges.  The  growth  on  this  first  or  original 
potato  will  be  quite  luxuriant,  and  the  individual  colonies  often 
difficult  to  recognize,  therefore  dilutions  are  made.  (Fig.  18.) 

From  the  original  or  first  slice,  a  small  portion  including  some 
of  the  meat  of  the  potato  is  spread  out  upon  the  surface  of  a 
second  slice,  which  is  first  dilution.  From  this  likewise  a  small 
bit  is  taken  and  spread  on  a  third  slice  or  second  dilution,  and 
here  usually  the  colonies  will  be  sparsely  enough  settled  to  study 
them  in  their  individuality. 

This  is  the  principle  carried  on  in  all  the  cultivations.  It  is 
a  physical  analysis. 


SOLID    TRANSPARENT    MEDIA. 
FIG.  18. 


47 


Method  of  inoculation.    (Woodhead  and  Hare.) 

Potato  and  Bread  Mash.  These  pastes  are  used  chiefly  in  the 
culture  of  moulds  and  yeasts.  Peeled  potatoes  are  mashed  with 
distilled  water  until  thick,  and  then  sterilized  in  flasks  f  of  an 
hour  for  three  successive  days. 

Bread  Mush. — Bread  devoid  of  crust,  dried  in  an  oven,  and 
then  pulverized  and  mixed  with  water  until  thick  and  sterilized 
as  above. 


CHAPTER  IX. 

SOLID  TRANSPARENT   MEDIA. 

Solid  Transparent  Media  are  materials  which  can  be  used  for 
microscopical  purposes  and  which  can  readily  be  converted 
into  liquids.  Such  are  the  gelatine  and  agar  materials.  % 

Gelatine.  Gelatine  is  obtained  from  bones  and  tendons,  and 
consists  chiefly  of  chondrin  and  gluten. 

The  French  golden  medal  brand  is  the  one  most  in  use,  found 
in  long  leaves  with  ribbed  lines  crossing  them. 

Koeh-Loffler  10  per  cent.  Bouillon-Gelatine.    To  the  meat- 
water  as  made  for  the  bouillon  are  added 
100  grammes  gelatine, 
10        "         peptone, 

5        "         salt, 
to  each  1000  grammes  of  the  meat-water. 


48 


ESSENTIALS  OP  BACTERIOLOGY. 


FIG,  19. 


This  is  placed  in  a  flask  and  gently  heated  until  the  gelatine 

is  dissolved. 

Neutralization  with  the  soda  and  then  cooking  in  water-bath 

for  1  hour  or  more  until  the  liquid  seems  clear,  then  add  white 

of  an  egg  and  boil  ^  hour 
longer ;  the  egg  will  produce 
a  clearer  solution  and  save 
much  trouble.  A  small  por- 
tion, while  hot,  is  now  filtered 
into  a  test-tube  and  tested  for 
alkalinity,  nnd  then  re-heated 
several  times,  watching  if  a 
cloudy  ppt.  forms. 

If  the  fluid  remains  clear 
upon  cooling,  the  remainder  of 
the  material  can  be  filtered, 
It  must  be  accomplished 
while  hot,  else  the  gelatine 
will  coagulate  and  prevent 
further  filtration. 

This  can  be  carried  on 
either  by  keeping  hot  the  so- 
lution continually  in  water- 
bath,  and  only  filtering  a  small 
quantity  at  a  time  through 
the  filter,  or  keeping  the  filter 
itself  hot,  either  with  a  hot 
water  filter  or  placing  the 
filter  in  steam  chest.  (Fig. 
19.) 
Clouding  of  Gelatine.  If  the  gelatine  does  not  come  out  clear, 

or  becomes  turbid  on  cooling,  it  may  be  due  to  several  things— 

1.  The  filter-paper  too  thin  or  impure. 

2.  Too  strongly  alkaline. 

3.  Cooked  too  long  or  not  Icng  enough. 

The  addition  of  the  white  of  an  egg,  as  before  mentioned,  will 
often  clear  it  up  ;  if  this  avails  not.  re-filtering  several  times,  and 
attention  to  the  few  points  mentioned. 


Hot-water  filter. 


SOLID    TRANSPARENT    MEDIA.  49 

Sterilizing  the  Gelatine.  The  gelatine  is  kept  in  little  flasks 
or  poured  at  once  into  sterile  test-tubes,  careful  not  to  wet  the 
neck  where  the  cotton  enters,  lest  when  cool  the  cotton  plug 
stick  to  the  tube. 

The  tubes  are  then  placed  in  steam-chest  for  three  successive 
days,  15  minutes  each  day  (or  in  water-bath  1  hour  a  day  for 
three  days).  Then  set  aside  in  a  temperature  of  15°  to  20°  C., 
and  if  no  germs  develop  and  the  gelatine  remains  clear,  it  can 
be  used  for  cultivation  purposes. 

Modifications.  The  amount  of  gelatine  added  to  the  meat- 
water  can  be  variously  altered,  and  instead  of  making  gelatine 
bouillon  the  gelatine  can  be  mixed  with  milk,  blood,  serum, 
urine,  and  agar-agar. 

The  nutrient  gelatine  bouillon  can  also  receive  additions  iu 
the  shape  of  glycerine  (4  per  cent,  to  6  per  cent,  being  added), 
or  reducing  agents  to  take  up  the  oxygen  present. 

Agar-Agar.  This  agent,  which  is  of  vegetable  origin,  derived 
from  sea-plants  gathered  on  the  coasts  of  India  and  Japan,  has 
many  of  the  properties  of  gelatine  ;  retaining  its  solidity  at  a 
much  higher  temperature,  it  becomes  liquid  at  90°  C.  and  con- 
geals again  at  45°  C.  Gelatine  will  liquefy  at  35°  C. 

It  is  not  affected  very  much  by  the  peptonizing  action  of 
the  bacteria. 

Preparation  of  Agar-Agar  Bouillon  or  Nutrient  Agar.  The 
ordinary  bouillon  is  first  made,  and  then  the  agar  cut  in  small 
pieces,  added  to  the  bouillon  (15  grammes  of  agar  to  1000 
grammes  bouillon). 

It  is  allowed  to  stand  several  minutes  until  the  agar  swells, 
and  then  placed  in  water-bath  or  steam-chest  for  six  hours  or 
more.  The  reaction  is  taken,  very  little  of  the  alkali  being 
sufficient  to  neutralize  it. 

A  white  of  an  egg  added,  and  boiled  for  several  hours  longer, 
when,  even  if  not  perfectly  clear,  it  is  filtered. 

The  filtering  process,  very  difficult  because  of  the  readiness 
with  which  the  agar  solidifies,  must  be  done  in  steam-chest  or 
with  hot-water  filter,  and  very  small  quantities  passed  through 
at  a  time,  changing  the  filter-paper  often. 
4 


50 


ESSENTIALS    OF    BACTERIOLOGY. 


Cotton  can  be  used  here  instead  of  filter-paper,  or  filtering 
entirely  dispensed  with  by  making  use  simply  of  decantation. 

As  Agar,  at  its  best,  is  seldom  clear,  a  little  more  or  less  opaque- 
ness, when  a  great  deal  of  trouble  and  time  can  be  saved 
thereby,  will  not  harm.  The  test-tubes  are  filled  as  with  the 
gelatine,  and  sterilized  in  the  same  manner.  AVhile  cooling, 
some  of  the  tubes  can  be  placed  in  a  slanting  position  so  as  to 
obtain  a  larger  surface  to  work  upon. 

Water  of  condensation  will  usually  separate  and  settle  at  the 
bottom,  or  a  little  white  sediment  remain  encysted  in  the  centre  ; 
this  cannot  easily  be  avoided,  nor  does  it  form  any  serious  obstacle. 
Glycerine  Agar.  The  addition  of  4  per  cent,  to  6  per  cent,  of 
glycerine  to  nutrient  agar  greatly  enhances  its  value  as  a  culture 
medium. 

Gelatine-Agar.    A  mixture  of  5  per  cent,  gelatine  and  0.75 
per  cent,  agar  combines  in  it  some  of  the  virtues  of  both  agents. 
Blood  Serum.    Blood  serum  being  rich  in  albumen  coagulates 
very  easily  at  70°  C.,  and  if  this  temperature  is  not  exceeded, 
a  transparent,  solid  substance  is  obtained 
upon  which  the  majority  of  bacteria  develop, 
and  some  with  preference. 

Preparation  of  Nutrient  Blood  Serum. 
If  the  slaughter  of  the  animal  can  be  super- 
vised, it  were  best  to  have  the  site  of  the 
wound  and  the  knife  sterilized  carefully, 
and  then  sterile  flasks  placed  to  receive  the 
blood  directly  as  it  flows. 

It  is  placed  on  ice  forty-eight  hours,  and 
then  the  serum  is  drawn  out  with  sterile 
pipettes  into  test-tubes ;  these  are  placed 
obliquely  in  an  oven  where  the  temperature 
can  be  controlled  and  maintained  at  a  cer- 
tain degree. 

Incubators  or  Brood-ovens.     Incubators 
or  brood-ovens,  as  such  ovens  are  called, 
consist  essentially  of  a  double-walled  zinc 
or  copper  chest,  the  space  between  the  walls  filled  with  water. 
The  oven  is  covered  with  some  impermeable  material  to  pre- 


FIG.  20. 


Flask  to  receive  blood 


SOLID    TRANSPARENT    MEDIA. 


51 


vent  the  action  of  surrounding  atmosphere.    (Fig.  22. )    It  is  sup- 
plied with  a  thermometer  arid  with  a  regulator.     The  regulator 

FIG.  21. 


Incubator  for  blood  serum. 

is  connected  with  the  Bunsen  burner,  and  keeps  the  thermometer 
at  a  certain  height. 

There  are  several  forms  of  regulators  in  use,  and  new  ones 
invented  continually. 

The  size  of  the  flame  is  regulated  by  the  expansion  ot  mer- 
cury, which,  as  it  rises,  lessens  the  opening  of  the  gas  supply. 
The  mercury  contracting  on  cooling  allows  more  gas  to  enter 
again.  (Fig.  23.) 

Koch  has  invented  a  safety  burner,  by  which  the  gas  supply  is 
shut  off  should  the  flame  accidentally  have  gone  out. 

Coagulation  of  Blood  Serum,  The  tubes  of  blood  serum 
having  been  placed  in  the  brood-oven,  are  kept  at  a  tempera- 


52 


ESSENTIALS  OF  BACTERIOLOGY. 


ture  of  65°  to  68°  C.,  until  coagulation  occurs  ;  then  removed 
and  sterilized. 


Fio.  22. 


Babe's  incubator. 


Thermo-regulators. 


Sterilization  of  Blood  Serum.  The  tubes  are  placed  3  to  4 
days  in  incubation  at  58°  C.,  and  those  tubes  which  show  any 
evidences  of  organic  growth  are  discarded. 

If  now,  at  the  end  of  a  week,  the  serum  remains  sterile  at  the 
ordinary  temperature  of  the  room,  it  can  be  used  for  experi- 
mental purposes. 

Perfectly  prepared  blood  serum  is  transparent,  of  a  gelatine- 
like  consistence,  and  straw-color.  It  will  not  liquefy  by  heat, 
though  bacteria  can  digest  it.  Water  of  condensation  always 
forms,  which  prevents  the  drying  of  the  serum. 

J31ood  serum,  formerly  much  more  used  than  now,  was  especi- 


INOCULATION  OF  GELATINE  AND  AGAR.     53 

ally  applicable  to  the  culture  of  tubercle  bacilli.  The  glycerine 
agar  has  now  superseded  it. 

Human  blood  serum  derived  from  placenta,  serum  from  ascitic 
fluid,  and  ovarian  cysts  are  prepared  in  a  similar  manner  to  the 
above. 

Other  Nutrient  Media.  Milk,  urine,  decoctions  of  various 
fruits  and  plants,  and  lately  for  cultivating  anaerobic  bacteria, 
eggs. 

Fresh  Egg  Cultures,  after  Hiippe.  The  eggs  in  the  shell  are 
carefully  cleaned,  washed  with  sublimate,  and  dried  with  cotton. 

The  inoculation  occurs  through  a  very  fine  opening  made  in 
the  shell  with  a  hot  platinum  needle  ;  after  inoculation,  the 
opening  is  covered  with  a  piece  of  sterilized  paper,  and  collodion 
over  this. 


CHAPTER  X. 

INOCULATION  OF  GELATINE  AND   AGAR. 

Glass  Slide  Cultures.  Formerly  the  gelatine  was  poured  on 
little  glass  slides  such  as  are  used  for  microscopical  purposes, 
and  after  it  had  become  hard,  inoculated  in  separate  spots  as 
with  potatoes. 

Test  Tube  Cultures.  The  gelatine,  agar,  or  blood  serum  having 
solidified  in  an  oblique  position,  is  smeared  on  the  surface  with 
the  material  and  the  growth  occurs,  or  the  medium  is  punctured 
with  a  stab  of  the  platinum  rod  containing  the  material.  The 
first  is  called  a  stroke  or  smear  culture,  the  second  a  stab  or  thrust 
culture.  In  removing  the  cotton  plugs  from  the  sterile  tubes  to 
carry  out  the  inoculation,  the  plugs  should  remain  between  the 
fingers  in  such  a  way  that  the  part  which  comes  in  contact  with 
the  mouth  of  the  tube  will  not  touch  anything. 

After  the  needle  has  been  withdrawn  the  plugs  c^re  re-inserted 
and  the  tubes  labelled  with  the  kind  and  date  of  culture. 

Plate  Cultures.  Several  tubes  of  the  culture  medium  are 
made  liquid  by  heating  in  water  bath,  and  then  inoculated  with 
the  material  as  follows  :  — 


64        ESSENTIALS  OF  BACTERIOLOGY. 

The  first  tube  is  called  original.  From  this  three  drops  (taken 
with  the  looped  platinum  rod)  are  placed  in  a  second  tube,  the 
rod  being  shaken  somewhat  in  the  gelatine  or  agar ;  this  is 
labeled  first  dilution  (a  colored  pencil  is  useful  for  such  markings). 

FIG.  24. 


Manner  of  holding  tubes  for  inoculation :  a,  tube  with  material ;  b,  tube  to  be 
inoculated ;  c,  cotton  plugs. 

From  the  first  dilution  three  drops  are  taken  into  a  third  tube, 
which  becomes  the  second  dilution. 

The  plugs  of  cotton  must  be  replaced  after  each  inoculation, 
and  during  the  same  must  be  carefully  protected  from  contami- 
nation. 


INOCULATION    OF    GELATINE    AGUE. 


55 


To  hasten  the  procedure  and  lessen  the  danger  of  contamina- 
tion, the  tubes  can  be  held  in  one  hand  aside  of  each  other,  each 

FIG.  25. 


FIG.  26. 


Manner  of  holding  plugs. 

plug  opposite  its  tube.     They  are  now  read}'  for  spreading  on 
glass  plates. 

Glass  Plates.  The  larger  the  surface  over  which  the  nutrient 
medium  is  spread  the  more  isolated  will 
the  colonies  be  ;  window  glass  cut  jn  rec- 
tangular plates  0x4  inches  in  size  is  used  ; 
about  ten  such  plates  are  cleaned  with  dry 
towel  and  placed  in  a  small  iron  box  or 
wrapped  in  paper ;  and  sterilized  in  the 
hot-air  oven  at  a  temperature  of  150°  C. 
for  ten  minutes.  (Fig.  20.)  When  the 
plates  have  cooled  they  are  placed  upon 
an  apparatus  designed  to  cool  and  so- 
lidify the  liquid  media,  which  is  now 
poured  upon  the  plates  from  the  inocu- 
lated test-tubes. 


A 


Iron  box  for  glass  plates. 


Nivellier  Leveling  and  Cooling  Apparatus.  Ice  and  water 
are  placed  in  a  shallow  round  glass  tray  ;  on  top  of  this  a  square 
plate  of  glass,  upon  which  the  culture  plate  is  placed,  and  cov- 
ering this  a  bell-glass. 

The  whole  is  upon  a  low,  wooden  tripod,  the  feet  of  which 
can  be  raised  or  lowered,  and  a  little  spirit-level  used  to  adjust 
it.  (Fig.  27.)  The  glass  plate  taken  out  of  the  iron  box  is  placed 
under  the  bell-glass.  The  tube  containing  the  gelatine  is  held 


56        ESSENTIALS  OF  B ACTERrOLOGY. 

in  the  (lame  a  second  to  singe  the  cotton  plug,  and  the  plug 
removed,  the  edges  of  the  tube  again  flamed,  the  bell-glas.s 
lifted,  and  the  inoculated  gelatine  carefully  poured  on  the  plate, 
leaving  about  one-third  inch  margin  from  the  borders  ;  the  lips 

FIG.  27. 


Nivellicr  leveling  and  cooling  apparatus. 

of  the  tube  being  sterile  can  be  used  to  spread  the  media  evenly. 
If  the  plate  is  at  all  cool,  the  fluid  will  solidify  as  it  is  being 
spread.  The  glass  cover  is  replaced  until  the 'gelatine  or  agar 
is  quite  solid  to  prevent  contamination. 

FIG.  28. 


Moist  chamber  with  plates  on  benches. 

When  the  gelatine  is  congealed,  the  plate  is  placed  upon  a 
little  glass  bench  or  stand  in  the  moist  chamber. 

The  Moist  CJmmbcr  Prepared  Out  of  Two  Glass  Dishes,  as  for 
tJie  Potato-  Culture*.  Then  glass  benches  are  so  arranged  that 
one  stands  upon  the  other.  In  order  to  avoid  confusion,  a  slip 
of  paper  with  a  number  written  on  it  is  placed  on  the  bench  be- 
neath each  plate.  As  the  original  or  first  plate  would  have  the 
colonies  developed  in  greatest  profusion,  it  is  placed  the  first 


INOCULATION  OF  GELATINE  AND  AGAR.     57 

day  on  the  topmost  bench  ;  but,  since  the  colonies  would  be 
likely  to  overrun  the  plate  and  allow  the  gelatine  to  drop  on  the 
lower  plates,  it  is  best,  as  soon  as  evidences  of  growth  appear, 
to  place  it  below,  and  watch  the  third  plate  or  second  dilution 
for  the  characteristic  colonies,  forgetting  not  all  this  time  to 
change  the  numbers  accordingly. 

The  date  of  culture  and  the  name  can  be  written  upon  the 
moist  chamber. 

Petri  Saucers,    Agar  hardens  very  quickly,  even  without  any 
especial  means  for  cooling,  and  it  does 
not  adhere  very  well  to  the  glass.     There-  FIG*  29. 

fore  it  is  better  to  follow  the  method  of 
Petri  and  use  little  shallow  glass  dishes, 
one  covering  the  other.  They  are  first 
sterilized  by  dry  heat,  and  then  the  in- 
oculated gelatine  or  agar  is  poured  into  Petri  saucerg 
the  lower  dish,  covered  by  the  larger  one, 

and  placed  in  some  cool  place,  different  saucers  being  used  for 
each  dilution. 

This  method  is  ver}'  useful  for  transportation  ;  the  saucers  can 
be  viewed  under  microscope  similar  to  the  glass  plates. 

Esmarch's  Tubes,  or  Rolled  Cultures.  This  method,  especially 
used  in  the  culture  of  anaerobic  germs,  consists  in  spreading  the 
inoculated  gelatine  upon  the  inner  walls  of  the  test  tube  in 
which  it  is  contained  and  allowing  it  to  congeal.  The  colonies 
then  develop  upon  the  sides  of  the  tube  without  the  aid  of 
other  apparatus.  The  method  is  useful  whenever  a  very  quick 
and  easy  way  is  required.  The  rolling  of  the  tube  is  done  under 
ice-water  or  running  water  from  the  faucet.  The  tube  is  held 
a  little  slanting,  so  as  to  avoid  getting  too  much  gelatine  around 
the  cotton  plug. 

The  tubes  can  be  placed  directly  under  the  microscope  for 
further  examination  -of  the  colonies. 

NOTE.— The  peptone  nutrient  gelatine,  blood  serum  and  agar 
can  now  be  purchased  already  prepared,  thus  saving  a  great 
deal  of  time  and  making  unnecessary  the  purchase  of  consider- 
able apparatus. 


58 


ESSENTIALS  OF  BACTERIOLOGY. 


CHAPTER  XI. 


THE   GROWTH    AND   APPEARANCES   OF   COLONIES. 


FIG.  30. 


Naked  eye  appearances  of  colonies. 


Macroscopic.    Depending  greatly  upon  the  temperature  of 
the  room,  which  should  be  about 
65°  C.,  the  colonies  develop  so  as 
to  be  visible  to  the  naked  eye  in 
two  to  four  days.     Some   require 
ten  to  fourteen  days,  and   others 
grow  rapidly,  covering    the  third 
dilution  in  thirty-six  hours,     The 
plate  should  be  looked  at  each  day. 
The  colonies  present  various  ap- 
pearances, from  that  of  a  small  dot, 
like  a  fly  speck,  to  that  resembling 
a  small  leaf.     Some  are  elevated, 
some  depressed,  and  some,  like  cholera,  cup-shaped,  umbilicated. 
Then  they  are  variously  pigmented.     Some  liquefy  the  gela- 
tine speedily,  others  not  at  all.     The  appearances  of  a  few  are 
so  characteristic  as  to  be  recognized  at  a  glance. 

Microscopic.  We  use  a  low-power  lens,  with  the  abbe  nearly 
shut  out,  that  is  the  narrowest  blender.  The  stage  of  the 
microscope  should  be  of  such  size  as  to  carry  a  culture  plate 
easily  upon  it. 

The  second  dilution  or  third  plate  is  usually  made  use  of,  that 
one  containing  the  colonies  sufficiently  isolated. 

These  isolated  ones  should  be  sought  for,  and  their  appearances 
well  noticed. 

There  may  be  two  or  three  forms  from  the  same  germ,  the 
difference  due  to  the  greater  or  less  amount  of  oxygen  that  they 
have  received,  or  the  greater  or  less  amount  of  space  that  they 
have  had  to  develop  in. 

The  microscopic  picture  varies  greatly  ;  now  it  is  like  the 
gnarled  roots  of  a  tree,  and  now  like  bits  of  frosted  glass  ;  the 
pictures  are  very  characteristic,  and  the  majority  of  bacteria 
can  be  told  thereby.  (Fig.  31.) 


GROWTH    AND    APPEARANCES    OF    COLONIES.        59 

Impression  or  "Klatseh"  Preparations.  In  order  to  more 
thoroughly  study  a  certain  colony  and  to  make  a  permanent 
specimen  of  the  same,  we  press  a  clean  cover-glass  upon  the 
particular  colony,  and  it  adheres  to  the  glass.  It  can  then  be 


FIG.  31. 


FIG.  32. 


Microscopic  appearances 
of  colonies. 


Klatsch  preparations. 


stained  or  examined  so.  The  Germans  give  the  name  of 
"Klatsch"  to  such  preparations.  Many  beautiful  pictures  can 
be  so  obtained. 

Fishing.  To  obtain  and  examine  the  individual  members  of 
a  particular  colony  the  process  of  fishing,  as  it  is  called,  is 
resorted  to. 

The  colony  having  been  placed  under  the  field  of  the  micro- 
scope, a  long  platinum  needle,  the  point  slightly  bent,  is  passed 
between  the  lens  and  the  plate  so  as  to  be  visible  through  the 
microscope,  then  turned  downward  until  the  colony  is  seen  to 
be  disturbed,  and  the  needle  is  dipped  into  the  colony.  This 
procedure  must  be  carefully  done,  lest  a  different  colony  be 
disturbed  than  the  one  looked  at,  and  an  unknown  or  unwanted 
germ  obtained. 

After  the  needle  has  entered  the  particular  colony,  it  is  with- 
drawn, and  the  material  thus  obtained  is  further  examined  by 
staining  and  animal  experimentation.  The  bacteria  are  then 
again  cultivated  by  inoculating  fresh  gelatine,  making  stab  and 
stroke  cultures. 


60        ESSENTIALS  OP  BACTERIOLOGY. 

It  is  necessary  to  transfer  the  bacteria  to  fresh  gelatine  about 
every  six  weeks,  lest  the  products  of  growth  and  decay  given 
oft*  by  the  organisms  destroy  them. 


CHAPTER  XII. 

CULTIVATION   OF   ANAEROBIC   BACTERIA. 

SPECIAL-  methods  are  necessary  for  the  culture  of  the  anaerobic 
variety  of  bacteria  in  order  to  procure  a  space  devoid  of  0x3* gen. 
FIG  33  Several  measures  have  been  adopted  of  which  the 
easiest  and  most  serviceable  are  the  following  : — 

Liborius's  High  Cultures.  The  tube  is  filled  about 
f  full  with  gelatine,  which  is  then  steamed  in  a  water 
bath  and  allowed  to  cool  to  40°  C.,  when  it  is  inocu- 
lated by  means  of  a  long  platinum  rod  with  small 
loop,  the  movement  being  a  rotary  vertical  one,  and 
the  rod  going  to  the  bottom  of  the  tube. 

The  gelatine  is  next  quickly  solidified  under  ice  ; 
very  little  air  is  present.  The  ana?robic  germs  will 
grow7  from  the  bottom  upward,  and  any  rcrobins 
present  will  develop  first  on  top,  this  method  being 
one  of  isolation. 

From  the  anaerobic  germs  grown  in  the  lower  part, 
a  stab  culture  is  made  into  another  tube  containing 
£  gelatine,  the  material  being  obtained  by  breaking 
test-tube  with  the  culture. 

Hesse's  Method.  A  stab  culture  having  been  made 
with  ana?robic  germs,  gelatine  in  a  semi-solid  condi- 
Liborius's  tjon  {s  p(nire(i  jnto  the  tube  until  it  is  full,  thus  dis- 

IIU'tlllMl.  -         .  -  .  T,.  _  .    . 

placing  the  air.     (r  ig.  .54. ) 

Esmarch's  Method.  Having  inoculated  a  tube  with  the 
microbe  the  gelatine  is  rolled  out  on  the  walls  of  the  tube,  a 
"roll  culture,"  and  the  rest  of  the  interior  filled  up  with  gela- 
tine, the  tube  being  held  in  ice  water  in  the  meanwhile.  The 
colonies  develop  upon  the  sides  of  the  tube  and  can  be  easily 
examined  microscopically. 


CULTIVATION    OF    ANAEROBIC    BACTERIA. 


61 


Gases  like  Hydrogen  to  replace  the  Oxygen.  Several  arra nge- 
ments  for  passing  a  stream  of  hydrogen  through  the  culture  : — 

Frankel  puts  in  the  test  tube,  a  rubber  cork  containing  tw° 
glass  tubes,  one  reaching  to  the  bottom  and  connected  with  a 
hydrogen  apparatus,  the  other  very  short,  both  bent  at  right 
angles.  When  the  hydrogen  has  passed  through  ten  to  thirty 
minutes,  the  short  tube  is  annealed  and  then  the  one  in  connec- 
tion with  the  hydrogen  bottle,  and  the  gelatine  rolled  out  upon 
the  walls  of  the  tube.  (Fig.  35.)  Hiippe  uses  eggs  as  described 
in  Chapter  IX. 


FIG.  34. 


FIG.  35. 


FIG.  3G. 


Hesse's  method. 


Frankel's  method. 


Buchner's  method. 


Use  of  JSrobic  Bacteria  to  remove  the  Oxygen.  Roux  inocu- 
lates an  agar  tube  through  a  needle  thrust  after  which  semi- 
solid  gelatine  is  poured  in  on  top.  When  the  gelatine  has  solidi- 
fied, the  surface  is  inoculated  with  a  small  quantity  of  bacillus 


62         ESSENTIALS  OF  BACTERIOLOGY. 

subtilis  or  some  other  aerobic  germ.     The  subtilis  does  not  allow 
the  oxygen  to  pass  by,  appropriating  it  to  itself. 

Buchner's  Method.  The  test  tube  containing  the  culture 
is  placed  within  a  larger  tube,  the  lower  part  of  which  con- 
tains an  alkaline  solution  of  pyrogallic  acid  or  some  other  agent 
which  absorbs  oxygen.  The  tube  is  then  closed  with  a  rubber 
stopper.  (Fig.  37.) 


CHAPTER  XIII. 

THE  WAY  IN  WHICH  BACTERIA  AFFECT  THE  ANIMAL  ORGANISM. 

BACTERIA  affect  the  animal  organism  by  depriving  the  cells 
of  the  body  of  oxygen  and  nitrogen  which  they  appropriate  to 
themselves  for  their  maintenance. 

They  do  more  than  this,  however,  for  in  their  secretions  and 
excretions  the  main  potency  lies. 

Ptomaines,  or  Cadaveric  alkaloids,  was  the  name  first  applied 
to  those  bodies  formed  during  putrefaction,  but  now  used  for 
all  alkaloids  or  bodies  of  a  basic  nature  formed  by  bacteria. 
Many  of  these  ptomaines  when  introduced  into  the  body  give 
rise  to  the  same  set  of  symptoms  as  the  bacteria  themselves  do, 
so  that  we  may  say,  bacteria  affect  the  animal  body  chiefly  through 
certain  toxic  principles  which  they  produce  and  which  can  be  isolated. 

Toxines  and  Toxalbumens.  Late  researches  claim  two  classes 
of  products  for  bacteria— the  one  toxic  and  destroyed  by  heat ; 
the  other  anti-toxic,  having  a  direct  action  upon  the  tissue  and 
preventing  further  infection.  Then  proteids  or  toxalbumens, 
products  extracted  from  pure  cultures,  which,  like  ptomaines, 
produce  symptoms  similar  to  those  of  the  bacteria  itself.  They 
are  amorphous  and  have  no  basic  action,  giving,  however,  all 
the  reactions  of  a  proteid  or  albumen. 

Filtration  of  Cultures.  These  products  are  isolated  from  the 
culture  after  the  bacteria  themselves  have  been  separated. 

A  filter  consisting  of  a  cylinder  of  porcelain,  asbestos  or 
kaolin,  through  which  the  culture  fluid  passes,  the  bacteria 
remainin^  behind,  is  called  the  Pasteur-Chamberland  filter. 


THE    ANIMAL    ORGANISM. 
FIG.  37. 


63 


Pasteur-Chamberland  filter  with  pressure. 
A.  Container.    H.  Filter.    K.  Porcelain.    P.  Air-pump. 


64        ESSENTIALS  OF  BACTERIOLOGY. 

(Fig.  37.)  The  culture  can  be  forced  through  or  allowed  to  filter 
slowly. 

The  gerraless  liquor  is  then  treated  with  various  agents, 
alcohol  and  acetic  acid  being  that  used  for  the  loxalbumen  of 
diphtheria,  and  a  white  amorphous  powder  is  at  length  obtained. 
These  agents  have  different  effects  in  different  doses,  and  are 
used  also  to  establish  an  immunity. 

Toxic  Bacteria.  Those  bacteria  which  produce  toxic  agents 
outside  of  the  body,  and  will  not  develop  in  the  body,  are 
called  toxic  bacteria.  They  are  pathogenic  only  in  the  sense 
that  their  products,  when  accidentally  introduced  into  the  body, 
cause  mischief. 

Infectious  Microbes.  Those  bacteria  which  can  develop  and 
do  develop  in  the  animal  body,  and  there  generate  products 
injurious  to  the  same,  are  called  infectious  bacteria,  or  pathogenic 
bacteria. 

The  Variations  of  Pathogenesis,  The  same  animals  under 
different  circumstances  can  be  differently  affected  by  the  same 
germ. 

The  ordinary  white  mouse  is  not  acted  upon  by  the  bacillus 
of  glanders. 

If,  however,  glycosuria  be  produced  in  the  mouse  in  any  way, 
it  speedily  becomes  attacked  by  the  bacillus. 

Different  animals  are  differently  affected  by  the  same  germs. 
As  we  said  before,  ordinarily  the  white  mouse  is  not  acted  upon 
by  the  bacillus  of  glanders,  but  the  house  mouse  is  at  all  times. 

The  bacterium  may  first  become  active  when  mixed  with 
certain  chemicals,  it  having  been  harmless  before. 

Attenuation  or  Weakened  Virulence.  Bacteria  can  be  les- 
sened in  action  either  temporarily  or  permanently,  or  made  in- 
active entirely  without  destroying  them.  There  are  the  natural 
decay  and  loss  of  strength  ;  and  successive  cultivation  in  artificial 
media  for  a  long  time  of  the  same  germ  also  destroys  its  potency. 
But  artificial  means  can  be  used,  such  as  the  use  of  chemical 
agents  added  to  the  nutrient  soil,  or  by  passing  the  germ  through 
animals  who  are  in  some  sense  immune,  and  are  less  affected  than 
the  animals  for  whom  it  is  strictly  pathogenic. 

Thus  the  bacillus  of  swine-erysipelas,  which  is  quite  virulent 


THE    ANIMAL    ORGANISM.  65 

for  pigs,  when  passed  through  rabbits  loses  much  of  its  power, 
and  again  introduced  into  pigs  will  sicken  them  but  slightly. 
Sunlight  or  any  other  agent  that  is  destructive  to  germs  will 
also  weaken  them  when  used  cautiously. 

Heat  is  the  surest  agent  to  lessen  the  action. 

The  longer  it  takes  to  produce  the  attenuation,  the  more  lasting  it  is. 

The  grade  of  virulence  will  oftentimes  remain  through  suc- 
cessive generations. 

Some  of  the  attenuations  have  been  named  according  to  the 
animal  that  they  will  affect ;  thus,  Mice-anthrax  is  a  culture  of 
anthrax  which  has  been  exposed  to  a  temperature  of  42.0  C.  for 
twenty  days,  and  which  will  destroy  nothing  larger  than  mice. 
A  culture  exposed  for  ten  days  will  kill  nothing  larger  than 
rabbits,  etc. 

The  only  explanation  that  can  be  given  of  attenuation  is  that 
the  microbes,  though  similar  in  appearance,  differ,  in  that  the 
weaker  ones  give  rise  to  less  toxic  products  ;  they  have  been 
exhausted. 

Nageli  makes  use  of  the  simili  of  the  sweet  and  bitter  almond, 
the  one  poisonous,  because  it  contains  amygdalin,  but  both  pos- 
sible to  be  borne  on  the  same  tree,  and  looking  alike  in  every 
particular. 

The  Resistance  of  the  Animal  Organism  to  Bacteria.  The 
body  is  in  some  sense  resistant  to  bacteria ;  to  some  more,  to 
others  less  ;  and  this  resistance  has  been  variously  explained. 

Chemical  Theory.  The  greater  or  less  alkalinityof  the  blood  di- 
minishing or  increasing  the  virulence  is  the  explanation  of  some. 

The  Theory  of  the  Action  of  the  Serum  of  the  Blood,  It  has 
been  lately  shown  that  the  serum  of  the  blood  has  a  direct  in- 
hibiting action  on  all  bacteria  ;  and  this  is  directly  dependent 
upon  the  quantity  and  quality  of  albumen  in  it.  It  was  for- 
merly thought  the  salts  of  the  blood  were  the  main  factors,  but 
these  only  serve  to  keep  the  albumen  in  good  condition. 

The  serum  will  hinder  the  growth  of  germs,  and  when  bac- 
teria are  injected  directly  into  the  blood,  they  soon  disappear. 
They  cannot  osmose  ;  they  collect  in  the  liver,  spleen,  and 
marrow  of  bones,  and  the  corpuscles  aid  growth.  The  serum 
of  different  animals  acts  differently  upon  the  same  microbe,  . 
5 


66        ESSENTIALS  OP  BACTERIOLOGY. 

Cellular  Theory  of  Metschinkoff,  The  phagocytes,  as  he 
terms  them,  are  auti-microbie.  They  are  the  soldiers  which 
endeavor  to  destroy  the  enemy.  If  the  cells  are  strong,  they 
become  the  victors ;  if  the  bacteria  are  stronger,  the  bacteria 
conquer  and  eat  up  the  cells. 

But  this  theory,  though  having  many  supporters,  is  opposed 
on  sufficient  grounds,  the  one  reason  being  that  whenever  cells 
become  the  residence  of  live  bacteria  they  suffer  ;  and  if  Mets- 
chinkoff  and  others  have  seen  bacteria  directly  enter  cells  and 
disappear,  it  is  that  they  were  destroyed  before,  and  that  the 

leucocytes  only  acted  as  scav- 
FlG'  38'  engers.     (Fig.  38.) 

And  the  late  researches 
with  the  serum  of  the  blood 
freed  of  its  cellular  elements, 
and  being  directly  anti-bac- 
teric,  would  seem  to  place  the 
phagocytes  in  the  background. 
The  matter  is,  however,  by 
no  means  settled. 

Phagocytes  of  Metschinkoff:  a.  Bacillus          m  ,  , , 

entering ;  6.  Bacillus  inclosed.  T°  Sum  UP.  We  have  P"th°- 

genic  microbes  such  as  give 

rise  to  products  injurious  to  the  animal  organism.  The  infec- 
tious ones  can  overcome  the  natural  resistance  of  the  animal 
body  and  develop  therein.  The  bacteria  can  have  this  activity 
lessened  or  destroyed  by  agents  which  are  injurious  to  their 
products,  so  as  to  render  them  inactive. 


CHAPTER  XIV. 

IMMUNITY. 

THE  natural  or  acquired  power  of  resistance  to  bacterial 
influences  is  called  immunity. 

Natural.  As  we  have  mentioned  before,  certain  animals  are 
naturally  not  acted  upon  by  bacteria  that  affect  other  animals. 
We  say  the  animal  or  person  is  immune  by  nature. 


IMMUNITY.  67 

Acquired.  But  immunity  can  be  acquired  by  various  means. 
We  know  that  one  case  of  smallpox  usually  protects  against 
other  attacks  and  so  with  morbilla  and  scarlatina.  This  is 
through  disease. 

Acclimatization  Immunity.  Various  diseases,  which  strangers 
to  a  climate  become  affected  with,  do  not  trouble  the  natives. 

Artificial  Immunity.    By  the  attenuated  virus,  as  with  anthrax. 

Inoculations  with  sterilized  cultures,  the  germs  being  destroyed. 
Even  certain  chemical  substances  when  injected  give  immunity 
from  certain  germs,  and  when  albumen  prepared  in  certain  ways 
was  injected,  immunity  also  obtained. 

The  various  theories  which  have  been  made  to  explain  the 
phenomena  of  immunity  are  all  unsatisfactory.  Some  say  that 
a  first  attack  destroys  the  agents  which  are  necessary  for  the 
disease  to  arise.  Others  say  that  certain  bacterial  products 
remain  in  the  body,  and  prevent  a  return  of  the  diseases — act  as 
guards.  Some  recently  claim  that  the  soil  is  rendered  unfit  for 
the  further  development  of  the  bacteria,  after  injection  of  some 
of  the  active  principles  of  the  bacterium.  Some  place  it  in 
the  blood  of  the  animals,  it  exerting  a  direct  germicidal  action. 

It  has  not  yet  been  satisfactorily  explained,  perhaps,  in  the 
late  activity  of  bacteriological  workers,  some  solution  of  this 
important  question  will  be  arrived  at. 

Cure  of  Infectious  Diseases  with  Bacteria  and  their  Products. 
Antagonism.  It  has  been  known,  and  is  easily  demonstrated, 
that  the  growth  of  one  bacterium  near  another  results  often  in 
the  destruction  of  one,  a  direct  antagonism  existing. 

Rabbits  suffering  with  anthrax  were  injected  with  large  quan- 
tities of  streptococcus  of  erysipelas  (pyogenes),  and  a  cure 
effected,  those  not  so  treated  dying. 

Several  other  diseases  have  been  so  treated  in  animals  with 
interesting  results. 

Toxalbumen  Injections.  When  diphtheria  is  produced  in 
animals,  an  injection  of  the  toxalbumen  of  diphtheria  will  cure 
the  same,  and  if  injected  first,  diphtheria  will  not  arise. 
Tetanus  has  been  cured  and  prevented  in  a  similar  manner. 

Tuberculin.  Dixon,  in  1889,  found,  by  injecting  products  of 
tuberculosis  cultures  in  glycerine,  that  Guinea-pigs  so  treated, 


68  ESSENTIALS    OF    BACTERIOLOGY. 

suffering  from  tuberculosis,  were  cured  ;  control  animals  dying. 
Koch,  in  1890,  applied  this  method  to  man,  and  the  final  results 
are  still  sub  juclice. 

Koch's  Rules  in  Regard  to  Bacterial  Cause  of  Disease, 
Before  a  microbe  can  be  said  to  be  the  cause  of  a  disease,  it 
must  — 

First.  Be  found  in  the  tissue  or  secretions  of  the  animal  suf- 
fering from,  or  dead  with  the  disease. 

Second.  It  must  be  cultivated  outside  of  the  body  on  artificial 
media. 

Third.  A  culture  so  obtained  must  produce  the  disease  in 
question  when  it  is  introduced  into  the  body  of  a  healthy 
animal. 

Fourth.  The  same  germ  must  then  again  be  found  in  the 
animal  so  inoculated. 


CHAPTER  XV. 

EXPERIMENTS   UPON    ANIMALS. 

THE  smaller  rodents  and  birds  are  the  ones  usually  employed 
for  inoculation,  as  rabbits,  Guinea-pigs,  rats  and  mice,  and 
pigeons,  and  chickens ;  sometimes  monkeys.  These  are  pre- 
ferred, because  easily  acted  upon  by  the  various  bacteria,  readily 
obtained,  and  not  expensive. 

The  white  mouse  is  very  prolific  and  easily  kept,  and  is  there- 
fore a  favorite  animal  for  experiment.  It  lives  well  upon  a  little 
moistened  bread.  A  small  box,  perforated  with  holes,  is  filled 
partly  with  sawdust,  and  in  this  ten  to  twelve  mice  can  be  kept. 
When  the  female  becomes  pregnant  she  should  be  removed  to 
a  glass  jar  until  the  young  have  opened  their  eyes,  because  the 
males,  which  have  not  been  raised  together,  are  apt  to  attack 
each  other. 

Guinea-pigs.  When  Guinea-pigs  have  plenty  of  light  and 
air  the}r  multiply  rapidly.  Therefore  it  is  best  to  have  them  in 
some  large  stall  or  inclosure.  They  can  be  fed  upon  all  sorts  of 
vegetables  and  grasses,  and  require  but  little  attention. 


EXPERIMENTS    UPON    ANIMALS.  69 

Methods  of  Inoculation,  Z  Inhalation.—  Imitating  the  natural 
infection,  either  by  loading  an  atmosphere  with  the  germs  in 
question  or  by  administering  them  with  a  spray. 

//.   Tlirougli  Skin  or  Mucous  Membrane. 

III.   With  the  Food. 

Method  of  Cutaneous  Inoculation.  The  ear  of  mice  is  best 
suited  for  this  procedure.  A  small  abrasion  made  with  the 
point  of  a  lancet  or  needle,  which  has  been  dipped  in  the  virus. 
The  animal  is  then  separated  from  the  rest  and  placed  in  a 
glass  jar,  which  is  partly  filled  with  sawdust  and  covered  with 
a  piece  of  wire-gauze. 

Subcutaneous.  The  root  of  the  tail  of  mice  is  used  for  this 
purpose.  The  hair  around  the  root  of  the  tail  is  clipped  off, 
and  with  a  pair  of  scissors  a  very  small  pocket  is  made  in  the 
subcutaneous  connective  tissue,  not  wounding  the  animal  any 
more  than  absolutely  necessary,  avoiding  much  blood.  The 
material  is  placed  upon  a  platinum  needle  and  introduced  into 
the  pocket,  solid  bodies,  with  a  forceps. 

To  hold  the  mouse  still  while  the  operation  is  going  on  a 
little  cone  made  of  metal  is  used.  The  mouse  just  fits  in  here. 
There  is  a  slit  along  the  top  in  which  the  tail  can  be  fastened, 
and  thus  the  animal  is  secure  and  immobile. 

Intravenous  Injections.  Rabbits  are  very  easily  injected 
through  the  veins.  Mice  are  too  small. 

The  ear  of  the  rabbit  is  usually  taken.  It  is  first  washed  with 
1-2000  bichloride,  which  not  only  disinfects,  but  also  makes  the 
vessels  appear  more  distinct.  The  base  of  the  ear  is  compressed 
to  swell  the  veins.  Then  a  syringe,  like  the  one  used  for  the 
injection  of  "  tubercnline,"  a  Koch  syringe,  which  can  be  easily 
sterilized,  is  filled  with  the  desired  amount  of  virus  and  slowly 
injected  into  any  one  of  the  more  prominent  veins  present. 
(Pig.  40.) 

Intra-peritoneal  Injection.  This  is  used  with  Guinea-pigs 
mostly.  The  abdominal  wall  is  pinched  up  through  its  entire 
thickness,  and  the  needle  of  the  syringe  thrust  directly  through, 
so  that  it  appears  on  the  other  side,  then  the  fold  let  go,  the 
needle  withdrawn  just  far  enough  so  as  to  be  within  the  cavity. 


70 


ESSENTIALS  OF  BACTERIOLOGY. 


Inoculation  in  the  Eye.  The  anterior  chamber  and  the  cornea 
are  the  two  places  used.  The  rabbit  is  fixed  upon  a  board  ; 
the  eyelids  held  apart  and  head  held  still  by  an  assistant.  A 
small  cut  is  made  in  the  cornea,  a  few  drops  of  cocaine  having 


FIG.  39. 


Manner  of  making  intravenous  injections  in  the  rabbit. 

first  been  introduced  in  the  eye.  The  material  is  passed 
through  the  opening  with  a  small  forceps,  and  with  a  few  strokes 
of  a  spoon  it  is  pushed  in  the  anterior  chamber. 

For  the  cornea  a  few  scratches  made  in  the  corneal  tissue 
will  suffice  ;  the  material  is  then  gently  rubbed  in. 

Inoculation  of  the  Cerebral  Membranes.  The  skin  and 
aponeurosis  cut  through  where  the  skull  is  the  thinnest.  Then 
the  bone  carefully  trephined,  and  the  dura  exposed.  In  Rabies 
inoculation,  the  syringe  containing  the  hydrophobia  virus  pierces 
the  dura  and  arachnoid,  and  the  virus  is  discharged  beneath  the 
latter. 

Intra-Tracheal.  The  bacteria  can  be  introduced  directly  into 
the  trachea,  thus  coming  in  contact  with  the  lungs. 

Intra-duodenal.— Cholera  germs  are  injected  into  the  intes- 
tines after  they  have  been  exposed,  by  carefully  opening  the 


EXPERIMENTS    UPON    ANIMALS.  71 

abdomen.  This  is  done  in  order  to  avoid  the  action  of  the 
gastric  juice. 

Obtaining  Material  from  Infected  Animals,  The  animal 
should  be  skinned,  or  the  hairs  plucked  out,  before  it  is  washed, 
at  least  the  portion  where  the  incision  is  to  be  made.  Then  the 
entire  body  is  washed  in  sublimate.  Two  sets  of  instruments 
are  required,  one  for  coarser  and  one  for  finer  work :  the  one 
sterilized  in  the  flame  ;  the  other,  to  prevent  being  damaged, 
heated  in  a  hot  air  oven. 

The  animal,  the  mouse  for  example,  is  stretched  upon  a  board, 
a  nail  or  pin  through  each  leg,  and  the  head  fixed  with  a  pin 
through  the  nose.  The  skin  is  dissected  away  from  the  belly 
without  exposing  the  intestines.  Then  the  ribs  being  laid  bare, 
the  sternum  is  lifted  up,  and  the  pericardium  exposed.  A  pla- 
tinum needle  dipped  into  the  heart  after  the  pericardium  has 
been  slit  will  give  sufficient  material  for  starting  a  culture.  If 
the  other  organs  are  to  be  examined,  further  dissection  is  made. 
If  the  intestines  were  first  to  be  looked  at,  they  would  be  laid 
bare  first. 

In  this  manner  material  is  obtained,  and  the  results  of  inocu- 
lation noted. 

Frequent  sterilization  of  the  instruments  is  desirable. 


PART  II. 
SPECIAL  BACTERIOLOGY. 


CHAPTER  I. 

NON-PATHOGENIC   BACTERIA. 

Special  Bacteriology.  Under  this  head  the  chief  character- 
istics of  individual  bacteria  will  be  detailed,  pathogenic  and  non- 
patkoycnic  being  the  main  divisions.  It  is  usual  to  describe  the 
non-pathogenic  first. 

Non-Pathogenic  Bacteria.  We  can  give  but  a  few  of  the  more 
important  varieties. 

Bacillus  Prodigiosus.  (Ehrenberg.)  This  bacillus,  formerly 
called  a  micrococcus,  is  very  common,  and  one  of  the  first 
noticed,  because  of  the  lively  red  color  it  forms  on  vegetables 
and  starchy  substances.  "  The  bleeding  host,"  miracles  being 
due  to  it. 

-F(/rrn.—  Short  rods,  often  in  filaments,  without  spores. 

Immobile. — Has  no  automatic  movements. 

Facultative  anaerobic,  that  is,  it  can  grow  without  air;  but 
the  pigment  requires  oxygen  to  show  itself. 

Growth.     Gelatine.     Liquefy  rapidly. 

Colonies. — At  first  white,  round  points  with  smooth  edge 
appearing  brown  under  microscope,  but  soon  changing  to  red. 

Stab  Cultures.— The  pigment  develops  on  the  surface,  the 
growth  occurring  all  along  the  line. 

Potato  is  well  suited  to  the  growth,  the  pigment  developing 
after  twelve  hours.  Agar  and  blood  serum  growths  do  well. 

Temperature.— Grows  best  at  25°  C. 

Varieties. — By  exposure  to  heat  of  brood-oven  during  several 
generations  the  power  to  produce  pigment  can  be  temporarily 
abolished. 

(72) 


NON-PATHOGENIC    BACTERIA.  73 

The  Pigment. — A  pigment-forming  body  is  created  by  the 
bacillus,  and  the  action  of  oxygen  upon  it  produces  the  color. 
It  is  insoluble  in  water,  slightly  soluble  in  alcohol  and  ether ; 
acids  fade  it,  alkalies  restore  the  color. 

Gases. — A  trimethylamin  odor  arises  from  all  culture. 

Stain. — Takes  all  aniline  dyes  easily  in  the  ordinary  way. 

Bacillus  Indicus.    (Koch.) 

Syn.  Micrococcus  Indicus.  Origin. — Found  in  the  stomach  of 
an  Indian  ape. 

Form. — Short  rods  with  rounded  ends.  No  spores.  Auto- 
matic movements  present ;  facultative  ancerobin. 

Growth.     Gelatine. — Liquefy  rapidly. 

Colonies. — Bound,  or  oval,  granular  margins ;  brilliant  red 
pigment. 

Stab  Cultures. — On  the  surface  the  pigment  shows  itself. 
Grows  well  on  other  media. 

Temperature'—  Grows  best  at  35°  C. 

Action  on  Animals.— In.  very  large  quantities,  if  injected  into 
the  blood,  a  severe  and  fatal  gastro- 
enteritis can  be  produced.  FlG-  40' 

Stain. — Takes  all  dyes. 

Bacillus  Mesentericus  Vulgatus. 

The  common   potato  bacillus  of 
Flugge. 

Habitat.  — Surface  of  the  soil,  on 
potatoes,  and  in  milk. 

Form. — Small    thick    rods    with 
rounded  ends,  often  in  pairs. 

Properties. — Very    motile  ;    pro- 
duce   abundant     spores;      liquefy     Colony  of  Bacillus  Mesentericus 
gelatine.  Vulgatus. 

Growth.— Rapid. 

Plate  .Colonies. — Round,  with  transparent  centre  at  first,  then 
becoming  opaque.  The  border  is  ciliated ;  little  projections 
evenly  arranged. 

Potato.— A  white  covering  at  first,  which  then  changes  to  a 
rough  brown  skin  ;  the  skin  can  be  detached  in  long  threads. 

Temperature.— Spores  at  ordinary  temperatures. 


74        ESSENTIALS  OF  BACTERIOLOGY. 

Spores.  —  Are  very  resistant ;  are  colored  in  the  manner  de- 
scribed in  first  part  of  the  book  for  spores  in  general. 

Bacillus  Megaterium  (de  Bury). 

Origin.— Found  on  cooked  cabbage. 

Form. — Large  rods,  four  times  as  long  as  they  are  broad, 
2.5  p.  Thick  rounded  ends.  Chains  with  ten  or  more  mem- 
bers often  formed. 

Properties.— Abundant  spore  formation  ;  very  slow  movement ; 
slowly  dissolves  gelatine. 

Growth.— Strongly  aerobic  ;  grows  quickly,  and  best,  at  a  tem- 
perature of  20°  C. 

Plate  Colonies.— Small,  round,  yellow  points  in  the  depth  of 
the  gelatine.  Under  microscope  irregular  masses. 

FIG.  41. 


Bacillus  Megaterium,  with  spores. 

Stab  Culture.— Funnel-shaped  from  above  downwards. 

Potato. — Thick  growths,  with  abundance  of  spores. 

Bacillus  Ramosus. 

Syn.  Bac.  Mycoides  (Fliigge).     Wurzel  or  root  bacillus. 

Origin.— In  the  upper  layers  of  garden  or  farm  grounds,  and 
in  water. 

Form.— Short  rods,  with  rounded  ends,  about  three  times 
as  long  as  they  are  thick  ;  often  in  long  threads  and  chains. 


NON-PATHOGENIC    BACTERIA.  75 

Properties. — Large,  shining,  oval  spores  ;  a  slight  movement ; 
liquefy  gelatine. 

Growth.— At  ordinary  temperatures,  with  plentiful  supply  of 
air. 

Plate  Colonies. — Look  like  roots  of  an  old  tree  gnarled  together, 
radiating  from  a  common  centre. 

Stab  Culture.— Soon  a  growth  occurs  along  the  needle  track, 
and  the  whole  resembles  a  pine  tree  turned  upside  down.  The 
gelatine  then  becomes  liquid,  a  thin  skin  floating  on  top,  and 
small  flakes  lying  at  the  bottom. 

Stroke  Culture. — Feathery  resemblance  is  produced. 

Staining. — Spores  stain  readily  with  the  ordinary  spore  stain. 

Bacterium  Zopfi.    (Kurth.) 

Origin. — Intestines  of  a  fowL 

Form. — Short  thick  rods  forming  long  threads  coiled  up, 
which  finally  break  up  into  spores,  which  were  once  thought  to 
be  micrococci. 

Properties. — Very  motile ;  does  not  dissolve  or  liquefy  gela- 
tine. 

Growth. — In  thirty  hours  abundant  growth;  aerobic;  grows 
best  at  20°  C. 

Plates.  —  Small  white  points  which  form  the  centre  of  a  very 
fine  netting.  With  high  power  this  netting  is  found  composed 
of  bacilli  in  coils,  like  braids  of  hair. 

Excellent  impress  or  "Klatsch"  preparations  are  obtained 
from  these  colonies. 

Staining.  —  Ordinary  dyes. 

Bacillus  Subtilis.     (Hay  Bacillus.)     Ehrenberg. 

Origin. — Hay  infusions  ;  found  also  in  air,  water,  soil,  faeces, 
and  putrefying  liquids. 

Form. — Large  rods,  three  times  as  long  as  broad ;  slight 
roundness  of  ends,  transparent ;  seldom  found  singly ;  usually 
in  long  threads.  Flagella  are  found  on  the  ends.  Spores  of 
oval  shape,  strongly  shining,  very  resistant. 

Properties. — Very  motile  ;  dissolves  gelatine. 

Growth. — Rapidly  ;  strongly  aerobic. 

Plate.— Round,  gray  colonies,  with  depressed  white  centre. 


76  ESSENTIALS    OF    BACTERIOLOGY. 

Under  microscope  the  centre  yellow ;  the  periphery  like  a  wreath, 
with  tiny  little  rays  projecting  ;  very  characteristic. 

Potato. — A  thick  moist  skin  forms  in  twenty-four  hours. 

Staining. — Rods,  ordinary  stain,  spores,  spore  stain. 

It  is  easily  obtained  by  covering  finely  cut  hay  with  distilled 
water,  and  boiling  a  quarter  of  an  hour.  Set  aside  forty-eight 
hours.  A  thick  scum  will  show  itself  on  the  surface  composed 
of  the  subtilis  bacilli. 

Bacillus  Spinosus.    (Luderitz.) 

Called  spinosus  because  small  spine-like  processes  are  formed 
by  the  colonies. 

Origin.— In  the  juices  of  the  body  of  a  mouse  and  guinea-pig 
which  were  inoculated  with  garden  earth. 

Form.— Large  rods,  straight,  some  slightly  bent,  ends  rounded  ; 
often  in  long  threads. 

Properties.  —  Large  spores,  the  bacillus  enlarging  to  allow  the 
spores  to  develop ;  very  motile  ;  gelatine  slowly  liquefied.  A 
gas  is  formed  in  the  culture  having  an  odor  like  Swiss  cheese. 

Grotcth. — The  growth  occurs  at  ordinary  temperatures  only 
when  the  oxygen  is  excluded.  Very  strongly  anaBrobic.  Glu- 
cose added  to  the  gelatine  (1  to  2  per  cent.)  increases  the  nutri- 
tive value. 

Colonies  in  roll  cultures  and  high  stab  cultures  appear  as  little 
spheres  surrounded  by  a  zone  of  liquefied  gelatine.  In  the 
deeper  growths  thorn-like  projections  or  spines  develop  pro- 
ceeding from  a  gray -colored  centre. 

Staining.—  With  ordinary  methods.  This  bacillus,  being 
strongly  anterobic,  must  be  cultured  with  the  usual  care  taken 
with  anserobins. 

Some  Bacteria  found  in  Milk.  Bacillus  Acidi  Lactici. 
(Iliippe.) 

Origin.— In  sour  milk. 

Form.— Short  thick  rods,  nearly  as  broad  as  they  are  long, 
usually  in  pairs. 

Properties. — Immotile.  Spores  large  shining  ones.  Do  not 
liquefy  gelatine.  Breaks  up  the  sugar  of  milk  into  lactic  acid 
and  carbonic  acid  gas,  the  casein  being  thereby  precipitated. 

Growth.— Slowly  ;  is  facultative  anaerobic.   Grows  first  at  10° C. 


NON-PATHOGENIC    BACTERIA.  77 

Plate  Colonies.— First  small  white  points,  which  soon  look  like 
porcelain,  glistening.  Under  microscope  the  surface  colonies 
resemble  leaves  spread  out. 

Stab  Culture. — A  thick  dry  crust  with  cracks  in  it  forms  on  the 
surface  after  a  couple  of  weeks. 

Attenuation. — If  cultured  through  successive  generations,  they 
lose  the  power  to  produce  fermentation.  Several  other  bacteria 
will  give  rise  to  lactic  acid  fermentation  ;  but  this  especial  one 
is  almost  constantly  found,  and  is  very  wide  spread. 

In  milk,  it  first  produces  acidity,  then  precipitation  of  casein, 
and  finally,  formation  of  gases. 

A  bacillus  described  by  Grotenfeldt,  and  called  Bacterium 
Acidi  Lactici,  forms  alcohol  in  the  milk.  It  was  found  in  milk 
in  Bavaria. 

Bacillus  Butyricus.    (Hiippe.) 

This  bacillus  causes  butyric  acid  fermentation. 

Origin. — Found  in  milk. 

.Form.  — Short  and  long  thin  rods  with  rounded  ends;  large 
oval  spores,  seldom  forming  threads. 

Properties. — Very  motile  ;  liquefies  gelatine  rapidly  ;  produces 
gases  resembling  butyric  acid  in  odor.  In  milk  it  coagulates  the 
casein,  decomposes  it,  forming  peptones  and  ammonia,  with  a 
bitter  taste,  and  butyric  acid  fermentation. 

Growth.— Quickly,  at  35°  to  40°  0.,  with  oxygen. 

Colonies.  Plate.  —  Small  yellow  points  which  soon  run  together, 
becoming  indistinguishable. 

Stab  Culture.  — A  small  yellow  skin  formed  on  the  surface  with 
delicate  wrinkles  ;  cloudy  masses  in  the  liquefied  portion. 

Staining. — With  ordinary  stains. 

Bacillus  Amylobacter  (Van  Tiegham) ;  or,  Clostridium  Buty- 

ricum.     (Prazmowsky.) 

Origin. — Found  in  putrefying  plant-infusions,  in  fossils,  and 
conifera  of  the  coal  period. 

Form, — Large,  thick  rods,  with  rounded  ends,  often  found  in 
chains.  A  large  glancing  spore  at  one  end,  the  bacillus  becoming 
spindle-shape  in  order  to  allow  the  spore  to  grow ;  hence  the 
name  clostridium. 


78        ESSENTIALS  OF  BACTERIOLOGY. 

FIG.  42.  Properties. — Very   motile  ;    gases    arise 

with  butyric  smell.  In  solutions  of  sugars, 
lactates  and  cellulose-containing  plants, 
and  vegetables,  it  gives  rise  to  decomposi- 
tions in  which  butyric  acid  is  often  formed. 
Casein  is  also  dissolved. 

Like  granulose,  a  watery  solution  of 
I  S  .-  h)  iodine  will  color  blue  some  portions  of  the 
1  >H  bacillus  ;  therefore  it  has  been  called  amy- 

(P    1     li     «         fofracter. 

\     «  Growth. — It  is  strongly  anicrebic,   and 

^      B|  j  has  not  yet  been  satisfactorily  cultivated. 

f  i  Bacillus  Lactis  Cyanogenus.    Bacterium 

Bacillus  Amyiobacter.      Syncyanum,  (Hiippe.) 

Origin.— Found  in  blue  milk. 

Form. — Small  narrow  rods  about  three  times  longer  than  they 
are  broad  ;  usually  found  in  pairs.  The  ends  are  rounded. 

Properties. — They  are  very  motile ;  do  not  liquefy  gelatine ; 
form  spores  usually  in  one  end.  A  bluish-gray  pigment  is  formed 
outside  of  the  cell,  around  the  medium.  The  less  alkaline  the 
media  the  deeper  the  color.  It  does  not  act  upon  the  milk  other- 
wise than  to  color  it  blue. 

Growth. — Grows  rapidly,  requiring  oxygen.  Colonies  on  plate. 
Depressed  centre  surrounded  by  ring  of  porcelain-like  bluish 
growth.  Dark  brown  appearance  under  microscope. 

Stab  Culture. — Grows  mainly  on  surface ;  a  nail-like  growth. 
The  surrounding  gelatine  becomes  colored  brown. 
Potato. — The  surface  covered  with  a  dirty  blue  scum. 
Attenuation. — After  prolonged  artificial  cultivation  loses  the 
power  to  produce  pigment. 

Staining. — By  ordinary  methods. 

Bacillus  Lactis  Erythrogenes.  Bacillus  of  Red  Milk.  (Hiippe 
and  Grotenfeldt.) 

Origin.— Found  in  red  milk,  and  in  the  faeces  of  a  child. 
Form. — Short  rods,  often  in  long  filaments,  without  spores. 
Properties.— Does  not  possess  self-movement.    Forms  a  nause- 
ating odor ;  liquefies  gelatine.   Produces  a  yellow  pigment  which 
can  be  seen  in  the  dark,  and  a  red  pigment  in  alkaline  media, 


NON-PATHOGENIC    BACTERIA.  79 

away  from  the  light.  In  milk  it  produces  the  yellow  cream  on 
top  of  the  blood-red  serum,  or,  fluid  in  the  centre,  and  at  the 
bottom  the  precipitated  casein. 

Growth. — Grows  rapidly  in  bouillon  and  on  potatoes ;  slower 
on  the  other  media  Plates.  A  cup-like  depression  in  the  centre 
of  the  colony,  with  a  pink  coloration  around  it,  the  colony  itself 
being  slightly  yellow. 

Stab  Culture. — The  growth  mostly  on  surface.  The  gelatine 
afterwards  colored  red  and  liquefied. 

Potato. — A  golden  yellow  pigment  formed  at  37°  C.,  after  six 
days. 

Some  Non-Pathogenic  Bacteria  found  in  Water.  The  bacteria 
found  here  are  very  often  given  to  producing  pigments  or  phos- 
phorescence, and  are  in  great  number.  The  more  common  ones 
only  will  be  described. 

Bacillus  Violaceus. 

Origin.  — Water. 

Form. — A  slender  rod  with  rounded  ends,  three  times  as  long 
as  it  is  broad,  often  in  threads  ;  middle-sized  spores. 

Properties. — Very  motile  ;  forms  a  violet-blue  pigment,  which 
is  soluble  in  alcohol,  and  depends  upon  oxygen  for  its  growth. 
Rapidly  liquefies  gelatine,  but  not  agar. 

Growth.— Grows  fairly  quick,  is  facultative  anaerobic. 

Cultures  on  Plate. — At  first  the  colonies  look  like  inclosed  air- 
bubbles.  Low  power  shows  irregular  masses,  with  a  centre 
containing  the  pigment  and  a  hairy-like  periphery. 

Stab  Culture. — Cone-like  liquefaction  containing  air,  and  the 
pigment,  in  separated  granules,  lying  towards  the  bottom. 

Stroke  Culture  on  Agar.— A  violet,  ink-like  covering  which 
remains  for  years. 

Bacillus  Coeruleus.    (Smith.) 

Origin. — Schuylkill  water. 

Form. — Very  thin  rods  ;  2.5  /*.  long,  0.5  /t*.  wide  ;  often  in 
threads  ;  spores  were  not  found. 

Properties. —  Liquefies  gelatine;  produces  a  very  deep-blue 
pigment. 

Growth. — Slowly,  with  oxygen,  at  ordinary  temperature. 

Plate. — Round  colonies  on  the  surface  of  bluish  color. 


80        ESSENTIALS  OF  BACTERIOLOGY. 

Stab  Cultures. — A  cup-shaped  liquefaction  along  the  needle 
thrust,  with  a  sparse  growth,  the  liquefied  portion  appearing 
blue. 

Fluorescent  Bacteria.    Several  kinds  present  in  water. 

Bacillus  Erythrosporus.    (Eidarn.) 

Origin.—  Drinking  water  and  putrefying  albuminous  solutions. 

Form. — Slender  rods  often  in  short  threads,  with  spores  of 
oval  shape,  and  appearing  as  if  stained  with  fuchsin. 

Properties. — Motile  ;  does  not  dissolve  gelatine  ;  produces  a 
greenish-fluorescent  pigment  which  appears  yellow  in  reflected 
light,  but  green  on  transmitted  light. 

Growth. — Somewhat  quickly  ;  facultative  anaerobic ;  growth 
only  at  ordinary  temperatures. 

Plates. —  White  colonies,  with  greenish-yellow  fluorescence 
around  each  colony.  Under  microscope  the  periphery  appears 
radiated. 

Stab  Cultures.— Good  growth  along  the  needle  thrust;  the 
whole  gelatine  gives  out  the  fluorescence. 

Bacillus  Fluorescens  Liquefaciens. 

Ch'iyin. — Water,  and  from  conjunctival  sac. 

Form. — Very  fine  little  rods  ;  no  spores. 

Properties. — Motile  ;  forms  a  greenish-yellow  fluorescent  pig- 
ment ;  liquefies  gelatine. 

Growth.— Rapid  only  at  ordinary  temperatures,  and  strongly 
aerobic. 

Plates. — Round  colonies,  cup-shaped  depressions,  the  solid 
gelatine  that  remains  becoming  colored  with  greenish-yellow 
fluorescence. 

Stab  Culture.— On  the  surface,  air-bubble  depressions ;  the 
white  colonies  in  the  bottom  of  these  depressions,  and  the  solid 
gelatine  around  the  inoculation  shining  with  the  fluorescence. 

Phosphorescent  Bacteria.  Six  varieties  of  phosphorescent 
bacteria  have  been  described  ;  they  are  found  usually  in  sea- 
water,  or  upon  objects  living  in  the  sea. 

Bacillus  Phosphorescens  Indicus.    (Fischer.) 

Origin. — Tropical  waters. 

Form.  —  Thick  rods,  with  rounded  ends,  sometimes  forming 
long  threads. 


NON-PATHOGENIC    BACTERIA.  81 

Properties. — Very  motile  ;  liquefying  gelatine  at  a  tempera- 
ture of  25°  to  30°  C.,  with  oxygen  and  a  little  moisture,  and  in 
the  dark,  a  peculiar  electric-blue  light  develops  a  phosphores- 
cence. 

Growth. — Slowly  ;  must  have  oxygen  ;  does  not  grow  under 
10°  C.  or  over  50°  C. 

Plates. — Little  round,  gray  points,  which  under  low  power 
appear  as  green  colonies  with  reddish  tinge  around  them. 
Cooked  fish,  when  smeared  upon  the  surface  with  a  little  of  the 
culture,  show  the  phosphorescence  most  marked.  Grows  well 
on  potatoes  and  blood-serum. 

Bacillus  Phosphorescens  Indigenus,    (Fischer.) 

Origin. — Waters  in  the  northern  part  of  Germany.  It  differs 
from  the  Indian  bacillus,  in  that  it  grows  at  a  temperature  of 
5°  C.,  and  does  not  develop  upon  potatoes  or  blood-serum. 

Bacillus  Phosphorescens  Gelidus.    (Fb'rster.) 

Origin. — Surfaces  of  salt-water  fish. 

Form. — Short,  thick  rods,  looking  oval  sometimes  ;  zoogkea 
are  often  formed. 

Properties. — Motile  ;  does  not  liquefy  gelatine ;  a  beautiful 
phosphorescence  from  the  surface  of  fish  ;  it  can  be  photographed 
by  its  own  light. 

Colonies. — Grows  best  between  0°  and  20°  C.  ;  grows  slowly, 
and  mostly  on  the  surface.  The  material  must  contain  salt. 
A  bouillon  made  with  sea-water,  or  3  to  4  per  cent,  common 
salt  will  suffice.  The  colonies  appear  as  those  of  the  Phospho- 
rescens Indicus. 

Fresh  herring  laid  between  two  plates  will  often  show  phos- 
phorescence in  twenty-four  hours. 

The  other  three  varieties  require  glucose  in  the  culture  before 
they  give  out  any  glow.  They  are  Bacterium  Pflugeri,  Bact. 
Fischeri,  and  Bact.  Balticum.  They  do  not  dissolve  gelatine. 

Several  very  indistinct  species,  found  in  waters  from  factories 
and  in  some  of  the  mineral  waters,  deserve  yet  to  be  men- 
tioned. They  have  been  given  various  names  by  observers ; 
almost  a  new  classification  created.  Such  are  the  crenothrix, 
dadothrix,  and  beggiatoa. 
6 


82        ESSENTIALS  OF  BACTERIOLOGY. 

Crenothrix  Kiihniana.  (Rabenhorst.)  Long  filaments  joined 
at  one  end  ;  little  rod-like  bodies  form  in  the  filaments ;  and 
these  break  up  into  spores. 

ZoogUea  are  also  formed  by  means  of  spores ;  and  these  can 
become  so  thick  as  to  plug  up  pipes  and  carriers  of  water. 
They  are  not  injurious  to  health. 

Cladothrix  Dichotoma.  (Cohn.)  Very  common  in  dirty 
waters.  The  filaments  branch  out  at  acute  angles,  otherwise 
resembling  the  crenothrix ;  accumulations  of  ochre-colored 
slime,  consisting  of  filaments  of  this  organism,  are  found  in 
springs  and  streams. 

Beggiatoa  Alba.  (Vancher.)  The  most  common  of  this 
species.  The  distinction  between  this  and  the  preceding  species 
lies  in  the  presence  of  sulphur  granules  contained  in  the  struc- 
ture, and  hence  they  are  often  found  where  sulphur  or  sulphides 
exist ;  but  where  the  remains  of  organic  life  are  decomposing 
they  can  also  be  found. 

Several  large  spirilla  and  vibrio  live  in  bog  and  rain-water, 
but  our  space  does  not  suffice  to  describe  them. 

Bacterium  Ureee. 

Origin. — Decomposed  ammoniacal  urine. 

Form. — Thick,  little  rods,  with  round  ends  one-half  as  thick 
as  they  are  long. 

Properties.—  Does  not  dissolve  gelatine ;  changes  urea  into 
carbonate  of  ammonia. 

Growth. — At  ordinary  temperatures,  very  slowly.  In  two  days 
on  gelatine  very  minute  points,  which  in  ten  days  have  the  size 
of  a  cent.  The  colonies  grow  in  concentric  layers. 

Micrococcus  Ureae.     (Pasteur  and  Van  Tieghain.) 

Orirjin. — Decomposed  urine  and  in  the  air. 

Form. — Cocci,  diplococci,  and  steptococci. 

Properties. — Decomposes  urea  into  carbonate  of  ammonia  ; 
does  not  liquefy  gelatine. 

Growth. — Grows  rapidly,  needing  oxygen ;  can  remain  sta- 
tionary below  0°  C. ;  growing  again,  when  a  higher  temperature 
is  reached. 

Colonies  on  Plate. — On  the  surface  like  a  drop  of  wax. 


NON-PATHOGENIC    BACTERIA.  83 

Stab  Cultures. — Looks  like  a  very  delicate  thread  along  the 
needle  thrust. 

Spirillum.    Spirillum  Rubrum.    (Esmarch.) 

Origin. — Body  of  a  mouse  dead  with  septicaemia. 

.Form.— Spirals  of  variable  length,  long  joints,  tlagella  on  each 
end  •,  no  spores. 

Properties. — Does  not  liquefy  gelatine  ;  very  motile  ;  produces 
a  wine-red  pigment,  which  develops  only  by  absence  of  oxygen. 

Growth. — Can  grow  with  oxygen,  but  is  then  colorless  ;  grows 
very  slowly  ;  ten  to  twelve  days  before  any  sign  ;  grows  best  at 
37°  C. 

Gelatine  Roll  Cultures.—  Small,  round;  first  gray,  then  wine- 
red  colonies. 

Stab  Cultures. — A  red-colored  growth  along  the  whole  line  ;  it 
is  deepest  below,  getting  paler  as  it  approaches  the  surface. 

Spirillum  Concentricum.    (Kitasato.) 

Origin. — Decomposed  blood. 

Form. — Short  spirals,  two  to  three  turns,  with  pointed  ends  ; 
it  has  flagella  on  the  ends. 

Properties. — Very  motile  ;  does  not  liquefy  gelatine. 

Growth.— Very  slow  ;  mostly  on  the  surface  ;  best  at  ordinary 
temperatures. 

Plates. — A  growth  of  rings  concentrically  arranged,  every 
alternate  one  being  transparent ;  the  furthest  one  from  the 
centre  possessing  small  projections. 

Stab  Cultures. — Growth  mostly  on  the  surface. 

Sarcina.  Cocci  in  cubes  or  packets  of  colonies.  A  great 
number  have  been  isolated  ;  many  producing  very  beautiful 
pigments.  The  majority  of  them  found  in  the  air. 

Sarcina  Lutea.    (Schroter.) 

Origin.  — Air. 

Form.— Very  large  cocci  in  pairs  ;  tetrads  and  groups  of 
tetrads. 

Properties. — Liquefies  gelatine  slowly  ;  produces  sulphur-yel- 
low pigment. 

Growth. — Slowly  ;  at  various  temperatures  ;  strongly  aerobic. 

Plates. — Small,  round,  yellow  colonies. 

Stab  Cultures.  —Grows  more  rapidly,  the  growth  being  nearly 


84        ESSENTIALS  OF  BACTERIOLOGY. 

all  on  the  surface,  a  few  separated  colonies  following  the  needle 
thrust  for  a  short  distance.  Agar,  a  very  beautiful  yellow, 
along  the  stroked  surface. 

Sarcina  Aurantica. — Flava,  rosea,  and  alba  are  some  of  the 
other  varieties.  Many  are  obtained  from  beer. 

Sarcina  Ventriculi.    (Goodsir.) 

Orifjin. — Stomach  of  man  and  animals. 

Form. — Colorless,  oval  cocci,  in  groups  of  eight  and  packets 
of  eight. 

Properties. — Does  not  liquefy  gelatine  ;  shows  the  reaction  of 
cellulose  to  iodine. 

Growth. — Rapid.  At  end  of  thirty-six  hours,  round,  yellow 
colonies,  from  which  colorless  cocci  and  cubes  are  obtained. 

Habitat. — They  are  found  in  many  diseases  of  the  stomach, 
especially  when  dilatation  exists. 


CHAPTER  II. 

PATHOGENIC  BACTERIA. 

WE  have  divided  this  part  into  two  portions.* 

I.  Those  bacteria  which  are  pathogenic  for  man  and  other 
animals. 

II.  Those  bacteria  which  do  not  affect  man.  but  are  patho- 
genic for  the  lower  animals. 

Here  again  it  will  only  be  possible  to  give  the  more  impor- 
tant bacteria  ;  there  are  many  diseases  in  which  micro-organisms 
have  been  found,  but  the}'  have  not  yet  been  proven  as  causa- 
tive of  the  disease,  and  have  also  been  found  in  other  diseases. 
We  cannot  treat  of  them  here. 

Bacillus  Anthracis.  (Bayer  and  Davaine.)—  Bayer  and  Da- 
vaine,  in  1850,  first  described  this  bacillus ;  but  Pasteur,  and 
later  -Kbc/t,  first  gave  it  the  importance  it  now  has. 

Synonyms. — Bacteridie  du  charbon  (Fr. ),  Milzbrand  bacillus 
(germ) ;  bacillus  of  splenic  fever,  or  malignant  pustule. 

Ori'jin.— In  blood  of  anthrax-suffering  animals. 


PATHOGENIC    BACTETUA.  85 

Form.—  "Rods  of  variable  length,  nearly  the  size  of  a  human 
blood-corpuscle,  broad  cup-shaped  ends ;  in  bouillon  cultures, 
long,  threads  are  formed,  with  large  oval  spores. 

FIG.  43. 


Anthrax  bacilli  in  human  blood  (fuchsin  staining),  Zeiss  1-12  oil  immersion. 
No.  4  ocular  taken  from  Vierordt. 

Properties. — Liquefies  gelatine  ;  immotile  ;  the  spores  are  very 
resisting,  living  twenty  years. 

Growth. — Grows  rapidly,  between  12°  C.  and  45°  C.,  and  re- 
quires plenty  of  oxygen,  not  growing  without  it ;  grows  well  in 
all  media. 

Plates  of  Gelatine. — Colonies  develop  in  two  days,  white  shiny 
spots,  which  appear  under  microscope  as  slightly  yellowish 
granular-twisted  balls,  like  a  ball  of  yarn  ;  each  separate  string 
or  hair,  if  looked  at  under  high  power,  being  composed  of  bac- 
teria in  line. 

Stab  Cultures. — A  white  growth  with  thorn-like  processes  along 
the  needle-track ;  later  on,  gelatine  liquefied,  and  fiaky  masses 
at  the  bottom. 

Potato.— A.  dry  creamy  layer,  and  when  placed  in  brood-oven, 
rich  in  spores. 


86 


ESSENTIALS  OF  BACTERIOLOGY 


Varieties.  Asporogenic. — By  cultivation  in  gelatine,  contain- 
ing 1  to  1000  ac-carbolic,  a  variety  develop  that  cannot  produce 
spores.  Also  involution  farms,  differing  from  the  usual  type. 


FIG.  44. 


FIG.  45. 


Stab  Cultures  of  Anthrax  in  Gelatine. 

Staining. — They  readily  take  all  the  aniline  dyes  with  the 
ordinary  methods.  To  bring  out  the  cup-shaped  concave  ex- 
tremities, a  very  weak  watery  solution  of  methylin  blue  is  best. 

Spores  are  stained  by  the  usual  method.  When  several  bacilli 
are  joined  together,  the  place  of  their  joining  looks  like  a  spore 
because  of  the  hollowed  ends.  The  double  staining  will  develop 
the  difference. 

Sections  of  tissue  are  stained  according  to  the  ordinary 
methods,  taking  Gram's  method  very  nicely. 

Pathoyeneste.— When  mice  are  inoculated  with  anthrax  mate- 
rial through  a  wound  in  the  skin,  they  die  in  twenty-four  hours 


PATHOGENIC    BACTERIA.  87 

from  an  active  septicaemia,  the  point  of  inoculation  remaining 
unchanged.  The  following  appearances  then  present  them- 
selves : — 

Peritoneum.— Covered  with  a  gelatinous  exudate. 

Spleen. — Very  much  swollen,  dark  red,  and  friable. 

Liver.  —  Parenchymatous  degeneration. 

Blood. — Dark  red.  The  bacilli  are  found  wherever  the  capil- 
laries are  spread  out,  in  the  spleen,  liver,  intestinal  villi,  and 
glomeruli  of  kidney,  and  in  the  blood  itself.  Only  when  the 
capillaries  burst  are  they  found  in  the  tubules  of  the  kidney. 

Mode  of  Entrance. — The  bacilli  can  be  inhaled,  and  then  a 
pneumonia  is  caused,  the  pulmonary  cells  containing  the  bacilli ; 
when  the  spores  are  inhaled,  a  general  infection  occurs. 

Feeding. — The  cattle  graze  upon  the  meadows,  where  the 
blood  of  anthrax  animals  has  flowed  and  become  dried,  the 
spores  remaining,  which  then  mix  with  the  grass  and  so  enter 
the  alimentary  tract ;  here  they  then  cause  the  intestinal  form 
of  the  disease,  ulcerating  through  the  villi. 

Local  Infection. — In  man  usually  only  a  local  action  occurs  ;  by 
reason  of  his  occupation — wool-sorter,  cattle-driver,  etc.,  he 
obtains  a  small  wound  on  the  hand,  and  local  gangrene  and 
necrosis  set  in. 

Pneumonia  by  inhalation  can  also  occur  in  man. 

Susceptibility  of  Animals.— Dogs,  birds,  and  cold-blooded  ani- 
mals affected  the  least ;  while  mice,  sheep,  and  guinea-pigs 
quickly  and  surely. 

Products  of  Anthrax  Bacilli. — A  basic  ptomaine  has  not  been 
found,  but  a  toxalbumen  or  proteid,  called  anthraxin,  has  been 
obtained.  A  certain  amount  of  acid  is  produced  by  the  virulent 
form,  alkali  by  the  weak. 

Attenuation  and  Immunity. — Cultures  left  several  days  at  a 
temperature  between  40°  and  42°  C.  soon  become  innocuous,  and 
when  injected  into  animals  protect  them  against  the  virulent 
form. 

Protective  Vaccination. — Animals  have  been  rendered  immune 
by  various  ways — by  inoculation  of  successive  attenuated  cul- 
tures ;  also  with  sterilized  cultures — that  is,  cultures  containing 
no  bacilli,  and  with  cultures  of  other  bacteria. 


88        ESSENTIALS  OF  BACTERIOLOGY. 

Habitat. — The  anthrax  disease  seems  confined  to  certain  dis- 
tricts in  Siberia,  Bavaria,  and  Auvergne,  and  mainly  during  the 
summer  months. 

The  bacillus  has  never  been  found  tree  in  nature. 

Bacillus  Tuberculosis.    (Koch.) 

This  very  important  bacillus  was  first  described,  demonstrated, 
and  cultivated  by  Koch,  who  made  his  investigations  public  on 

FIG.  4G. 


V 

O 


(   > 


s 

s 


^ 

X 


Tubercle  bacilli  in  sputum,  carbol-fuchsin,  and  methylin  blue.    Zeiss  1:12  oil 
immersion. 

the  24th  of  March,  before  the  Physiological  Society  of  Berlin, 
in  the  year  1882. 

Origin.— In  various  tubercular  products  of  man  and  other 
animals. 

Form.—  Very  slender  rods,  nearly  straight,  about  one-quarter 


PATHOGENIC    BACTERIA.  89 

the  size  of  a  red  corpuscle's  diameter,  their  ends  rounded,  usu- 
ally solitary,  often,  however,  lying  in  pairs  in  such  a  manner  as 
to  form  an  acute  angle.  Sometimes  they  are  '  S'-shaped.  In 
colored  preparations  little  oval  spaces  are  seen  in  the  rod,  which 
resemble  spores  ;  but  the  question  of  the  existence  of  spores  is 
still  undecided. 

Properties. — Does  not  possess  self-movement. 

Growt h.— Requires  special  media  for  its  growth,  and  a  temper- 
ature varying  but  slightly  from  37.5°  C.  It  grows  slowly,  de- 
veloping first  after  ten  days,  reaching  its  maximum  in  throe 
weeks.  It  is  facultative  anaerobic.  On  gelatine  it  does  not 
form  a  growth. 

Colonies  on  Mood  Serum.— Koch  first  used  blood  serum  for 
culture  ground,  and  obtained  thereon  very  good  growths.  Test- 
tubes  with  stroke  culture  were  placed  in  the  brood  oven  at  37°  C. 
for  ten  to  fourteen  days,  when  small  glistening  white  points  ap- 
peared which  then  coalesced  to  form  a  dry,  white,  scale-like 
growth.  Under  microscope  composed  of  many  fine  lines  con- 
taining the  tubercle  bacillus. 

Glycerine  Ayar.  —  J$y  adding  four  to  six  per  cent,  glycerine  to 
ordinary  agar-peptone  medium,  Nocard  and  Roux  obtained  a 
culture  ground  upon  which  tubercle  bacilli  grew  much  better 
than  upon  blood  serum.  This  is  now  almost  exclusively  used. 

Stroke  cultures  are  here  used  as  with  blood  serum.     They  are 
placed  in  brood-oven  after  inoculation,  and  remain  there  about 
ten  days,  at  a  temperature  of  37°  C. 
The   cotton   plugs   of   the   tubes   are  FIG.  47. 

covered  with  rubber  caps,  the  cotton 
first  having  been  passed  through  the 
flame,  and  moistened  with  a  few  drops 
of  sublimate  solution.  The  rubber 
cap  prevents  the  evaporation  of  the 
water  of  condensation  which  always 
forms,  and  keeps  the  culture  from  dry- 
ing up. 

The  growth  which  occurs  resembles 

the  ruga?  of  the  stomach,  and  some-  ^0 

times  looks  like  crumbs  of  bread  moist- 
ened. The  impression  or  "Klatsch"  preparation  shows  under 


90 


ESSENTIALS    OF    BACTERIOLOGY. 


the  microscope  a  thick  curled-up  centre  around  which  threads 
are  wound  in  all  directions.  And  these  fine  lines  show  the 
bacilli  in  profusion. 

FIG.  48. 


S  tf\     r~ 

,$\v^ 

$\ 


Klatsch  preparation. 

Potato.— It  can  be  cultivated  on  slices  of  potato  which  are 
placed  in  air-tight  test-tubes. 

Bouillon.  —  Bouillon  containing  four  per  cent,  glycerine  is  a 
very  good  nurture  ground. 

Varieties. — Dixon,  of  Philadelphia,  has  obtained  branched 
forms  of  bacilli  which  he  believes  to  be  degenerated  weakened 
ones. 

In  Sputum.— Little  granules  arranged  like  streptococci,  which 


PATHOGENIC    BACTERIA.  91 

take  the  characteristic  stain,  and  look  as  if  the  protoplasma  had 
been  destroyed  that  enclosed  them. 

FIG.  49. 


Growth  on  Agar. 

Bovine  tubercle-bacilli  are  about  one-third  smaller  than  human 
tubercle  bacilli. 

Staining. — The  tubercle  bacilli  require  special  methods  to 
stain  them,  and  a  great  number  have  been  introduced.  They 
are  stained  with  great  difficulty :  but  once  stained,  they  are 
very  resistant  to  decolorizing  agents.  Upon  these  facts  all  the 
methods  are  founded. 

It  will  only  be  necessary  to  describe  those  methods  principally 
in  use ;  and  as  the  examination  of  sputum  for  bacilli  is  of  so 
frequent  an  occurrence,  and  so  necessary,  it  is  well  to  detail  in 
particular  the  method  of  staining. 

Starting  with  the  sputum,  we  search  for  little  clumps  or  rolled- 
np  masses  ;  if  these  are  not  present,  the  most  solid  portions  of  the 
mucus  are  brought  with  forceps  upon  a  clean  cover-glass  ;  very 
little  suffices.  With  another  cover-glass  it  is  pressed  and  spread 
out  evenly  ;  drawing  one  glass  over  the  other,  we  obtain  two 
specimens,  and  these  put  aside  or  held  high  over  the  flame  until 
dry. 

If  we  desire  to  examine  the  specimen  quickly,  or  make  a 
hurried  diagnosis,  we  use  the  rapid  method,  with  hot  solutions  ; 
otherwise  we  let  it  stay,  in  cold  solution  until  the  next  morning 
the  advantages  of  which  will  be  later  on  described. 

TJie  Rapid  Method.— (B.  Frankel's  method  modified  by  Gab- 
bet.)  The  principle  is  to  combine  with  the  contrast  stain  the 


9          ESSENTIALS  OF  BACTERIOLOGY. 

decolorizing  agent ;  but  the  preparations  are  not  permanent ; 
the  method,  however,  is  very  useful. 

Two  solutions  are  required :  one  of  Ziehli's  carbol-fuchsin  ; 
the  other  Gabbet's  acid  methylin  blue.  (See  No.  X.,  on 
page  33.) 

The  cover-glass  containing  the  dried  sputum  is  passed  three 
times  through  the  flame,  as  described  in  the  general  directions. 
It  is  then  placed  in  the  carbol-fuchsin  solution  live  minutes  (cold), 
or  two  minutes  in  the  hot,  immediately  then  transferred  to  the 
second  solution,  the  acid  blue,  where  it  remains  one  minute, 
then  washing  in  water.  The  preparation  is  dried  between  filter- 
paper,  and  mounted  best  first  in  water.  Examined  with  oil- 
immersion. 

Aiwtlwr  Raitid  Method. — This  method  possesses  the  advan- 
tage of  giving  permanency  to  the  preparations  ;  but  the  bacilli 
are  distorted  and  ugly  crusts  form. 

Three  dishes  are  required  : — 

The  first  contains  nitric  acid  and  water  1-4. 

The  second  alcohol. 

The  third  distilled  water. 

They  are  arranged  one  after  the  other  in  the  above  order. 

Two  staining  solutions  must  be  at  hand,  carboi-fuchsin  and 
watery  methylin  blue.  The  cover-glass  containing  the  dried 
sputum  is  passed  three  times  through  the  flame,  then  covered 
with  a,  few  drops  of  carbol-fuchsin,  and  held  in  the  forceps  over 
the  flam?,  so  that  the  stain  will  boil  upon  the  glass.  With  a 
pipette  the  dye  is  constantly  added  and  kept  boiling  for  about 
one  minute.  It  is  then  decolorized  by  holding  it  in  the  first 
dish  until  it  appears  brownish-black,  then  directly  into  the 
second  dish,  when  the  alcohol  peals  off  the  red  color  in  little 
clouds,  and  it  becomes  nearly  colorless  ;  about  five  seconds 
suffice.  Then  it  is  placed  in  the  third  dish,  the  water  washing 
oft'  the  alcohol.  If  the  color  of  the  preparation  is  now  deeper 
than  a  slight  pink,  it  is  again  dipped  into  the  acid,  alcohol,  and 
back  into  the  water,  careful  not  to  hold  it  too  long  in  the  above 
solutions.  The  contrast  stain  is  now  applied,  a  few  drops  of  the 
methylin  blue  solution  left  on  cold  for  two  minutes  being  suffi- 
cient. The  glass  is  now  dried  and  mounted  on  the  slide  in 


PATHOGENIC    BACTERIA.  03 

Canada  balsam.    Examined  with  oil-immersion.     The  tubercle 
bacilli  red,  all  else  blue. 

Slaw  Method. — When  perfect  permanent  preparations  are 
desired  and  the  bacilli  to  be  seen  unaltered,  the  slow  method  is 
to  be  preferred,  and  it  is  to  be  recommended  whenever  the  time 
allows.  It  consists  simply  in  allowing  the  carbol-fuchsin  to  work 
upon  the  preparation  a  number  of  hours.  We  usually  place  the 
cover  glass  with  the  dried  sputum  and  which  has  been  drawn 
through  the  flame  three  times,  in  a  little  dish  containing  enough 
dye  to  allow  the  glass  to  be  immersed.  We  do  this  about  5  or  6 
o'clock  P.M.,  and  the  next  morning  the  preparation  is  ready 
for  decolorizing,  the  process  being  the  same  as  described  above, 
viz.,  25 %  nitric  acid,  alcohol  and  water,  and  the  contrast  stain 
methylin  blue.  We  thus  avoid  the  formation  of  ugly  crusts, 
the  bacilli  are  not  distorted,  the  specimen  is  permanent  and 
very  clear. 

Biederfs  Method  of  Collecting  Bacilli,  when  the  bacilli  are 
very  few  in  a  great  quantity  of  fluid,  as  urine,  pus,  abundant 
mucus,  etc.,  Biedert  advises  to  mix  15  c.cm.  of  the  fluid 
with  75  to  100  c.cm.  water  and  a  few  drops  of  potassium  or 
sodium  hydrate,  then  boiling  until  the  solution  is  quite  thin.  It 
is  placed  in  a  conical  glass  for  two  days,  and  bacilli  with  other 
morphological  elements  sink  to  the  bottom  of  the  glass ;  when 
(he  supernatant  liquid  is  decanted,  the  residue  can  be  easily 
examined.  In  this  way  bacilli  were  found  that  had  eluded 
detection  examined  in  the  ordinary  manner. 

Staining  Bacillus  Tuberculosis  in  Tissue  (sections}. — The  general 
method  of  Gram  can  be  used,  but  the  better  way  is  to  use  the 
following : — 

Carbol-fuchsin,  15  to  30  minutes. 

5  per  cent,  sulphuric  acid,  1  minute. 

Alcohol,  until  a  light-red  tinge  appears. 

Weak  methylin  blue,  3  to  5  minutes. 

Alcohol,  for  a  few  seconds. 

Oil  of  cloves,  until  cleared. 

Canada  balsam,  to  mount  in. 
Instead  of  carbol-fuchsin,  alcoholic  solution  offuchsin  or  aniline 


94        ESSENTIALS  OF  BACTERIOLOGY. 

water  fuchsin  can  be  used,  but  the  sections  must  remain  in  the 
stain  over  night. 

The  resisting  action  of  the  bacillus  to  acids  is  supposed  to 
be  due  to  a  peculiar  arrangement  of  the  albumen  and  cellulose 
of  the  cell  rather  than  to  any  particular  capsule  around  it. 

Patliogenesis. — When  a  guinea-pig  has  injected  into  its  peri- 
toneal cavity  some  of  the  diluted  sputum  containing  tubercle 
bacilli  it  perishes  in  about  three  weeks,  and  the  following 
picture  presents  itself  at  the  autopsy  :  at  the  point  of  inoculation 
a  local  tuberculosis  shmcs  itself,  little  tubercular  nodules  contain- 
ing the  characteristic  bacilli.  In  the  lungs  and  the  lymphatics, 
similar  tubercles  are  found,  a  general  tuberculosis. 

If  the  animal  lingers  a  few  weeks  longer,  the  tubercles  become 
necrosed  in  the  centre  and  degeneration  occurs,  the  periphery 
still  containing  active  bacilli,  cavities  having  formed  in  the 
centre. 

Since  the  bacilli  die  in  course  of  time,  killed  by  their  own  pro- 
ducts, their  number  forms  no  correct  guide  of  the  damage  present. 

Even  their  absence  in  the  sputum  does  not  preclude  ihe  ab- 
sence of  a  tubercular  process.  It  is  their  presence  only  that 
warrants  a  positive  declaration. 

They  are  found  in  the  blood  only  when  a  vessel  has  come  in 
direct  contact  with  a  tubercular  process  through  rupture  or 
otherwise.  They  have  been  found  in  other  secretions,  milk, 
urine,  etc. 

M«n  is  infected  as  follows  :  — 

Throutjh  wounds.—  Local  tuberculosis. 

Throwjk  nutrition. — Milk  of  tuberculosis  cows. 

Phthisical  patients  swallowing  their  own  sputum  and  causing 
an  intestinal  tuberculosis. 

Inhalation. — This  is  the  most  usual  way,  probably  constitu- 
ting the  cause  in  ,9(t  of  the  cases. 

The  sputum  of  phthisical  patients  expectorated  on  the  floors 
of  dwelling-houses  in  handkerchiefs,  etc.,  dries,  and  the  bacilli 
set  free  are  placed  in  motion  by  the  wind  or  rising  with  the  dust 
are  thus  inhaled  by  those  present.  When  the  sputum  is  kept 
from  drying  by  expectoration  in  vessels  containing  water,  this 
great  clanger  can  be  avoided. 


PATHOGENIC    BACTERIA.  95 

Nearly  all  the  cases  of  heredity  can  be  explained  in  this  man- 
ner. The  young  children,  possessing  very  little  resistance,  are 
constantly  exposed  to  the  infection  through  inhalation  and  also 
by  nutrition. 

Immunity. — No  one  can  be  said  to  be  immune,  though  per- 
sons who  have  been  greatly  weakened  would  offer  less  resistance 
than  health}''  individuals. 

Products  Of  Tubercle  Bacilli.  The  last  year  has  developed 
some  wonderful  facts  in  relation  to  this  important  deadly  bacillus. 
In  1889,  Dr.  Dixon,  then  Professor  of  Hygiene  at  the  Univer- 
sity of  Pennsylvania,  spoke  of  a  method  of  curing  tuberculosis 
in  guinea-pigs  and  with  products  obtained  from  the  bacillus ; 
not  much  was  thought  of  this  statement  at  the  time. 

In  August,  1890,  Koch,  before  the  Medical  Congress  claimed 
that  he  also  had  been  able  to  cure  tuberculosis  in  guinea-pigs, 
and  would  be  able  to  give  some  interesting  facts  later  on.  In 
November  he  claimed  that  he  had  obtained  reactions  in  man 
similar  to  those  in  the  guinea-pig,  and  believed  that  a  cure  was 
at  hand. 

In  the  excitement  which  followed  this  statement,  the  greatest 
hopes  were  raised  and  the  impossible  expected.  In  January, 
1891,  Koch  made  public  the  manner  of  preparing  the  lymph  or 
u  Tuberculin"  or  u  Kochin,"  as  it  was  variously  called  :  old  cul- 
tures of  tubercle  bacilli  mixed  with  60  per  cent,  glycerine  and 
filtered  through  a  Chamberlain-Pasteur  filter,  the  filtrate  thus 
obtained  being  a  dark-brown  liquid,  sp.  gr.  somewhat  higher  than 
water,  an  odor  like  "beef  extract,"  a  sweetish  taste,  not  soluble 
in  alcohol;  according  to  Jollas,  containing  50  per  cent,  water, 
and  showing  a  strong  Biuret  reaction  ;  he  thinks  it  therefore  a 
toxalhumen.  1  milligramme  of  the  lymph  is  supposed  to  contain 
but  ysnffo  milligramme  of  the  active  principle.  Dixon's  lymph 
is  obtained  in  a  very  similar  manner,  and  no  doubt  contains  the 
same  principle. 

Dixon  recommends  instead  of  using  the  pure  culture  for 
obtaining  the  lymph,  the  tuberculosis  lung  of  calf,  a  portion  of 
which  is  treated  with  water  and  glycerine,  and  then  filtered 
through  Chamberlain-Pasteur  filter  without  pressure. 

Manner  of  Usiwj  Koch's   Lymph.— One   milligramme   of  the 


96         ESSENTIALS  OF  BACTERIOLOGY. 

Koch's  lymph  is  injected  under  the  skin  of  one  suffering  with 
a  tubercular  process,  and  in  a  few  hours  to  a  few  days,  a  rise  of 
temperature,  tightness  about  the  chest,  and  exaggerated  cough- 
ing spells  take  place,  the  symptoms  varying  in  intensity; 
usually  a  secondary  rise  occurs  on  the  following  day.  The 
dose  has  been  gradually  increased  until  the  reactions  subsided, 
and  GOO  milligrammes  have  then  been  borne  without  any  reaction. 

On  Lupus  the  process  could  be  watched  and  was  very  char- 
acteristic ;  a  peculiar  redness  after  the  first  injection,  and  after 
a  few  more  injections  scabs  formed,  and  an  apparent  cure  seemed 
to  be  obtained,  but  relapses  were  common  and  but  very  few 
authentic  cures  if  any  can  now  be  had. 

Koch  believed  that  the  tuberculosis  tissue  was  rendered 
necrotic  by  this  toxic  principle,  making  the  soil  unfit  for  the 
bacilli  which  then  perished  or  were  expectorated. 

Virchow  dampened  the  excitement  and  ardor  by  showing  a 
great  diffusion  of  fresh  miliary  tubercles  in  the  bodies  of  persons 
who  had  died  and  who  had  been  treated  with  the  lymph.  Cool, 
careful,  and  untiring  study  and  time  taken  together  will,  we 
trust,  bring  a  happy  solution  and  a  genuine  remedy. 

Lepra  Bacillus.    ( H  a  n sen . ) 

Origin. — In  1880  Armauer  Hansen  declared,  as  the  result  of 
many  years'  investigation,  that  he  found  a  bacillus  in  all  leprous 
processes. 

Form. — Small  slender  rods  somewhat  shorter  than  tubercle 
bacilli,  otherwise  very  similar  in  appearance. 

In  the  interior  of  the  cell  two  to  three  oval  spaces  are  usually 
seen,  not  known  if  spores  or  otherwise. 

Properties. — They  are  immotile,  do  not  liquefy  the  nutrient 
media. 

Growth. — Bordoni-Uffreduzzi  have  obtained  growths  upon 
blood  serum  to  which  peptone  and  glycerine  had  been  added. 
The  growth  is  very  slow,  requiring  about  eight  days  at  a  tem- 
perature of  37°  C. 

Colonies. — Small  grayish  round  spots,  under  microscope  ap- 
pearing like  a  close-netted  spider  web  around  a  firm  centre. 

Stab  Cultures. — Show  a  waxy-like  growth  along  the  needle  track. 
/.— They  resist  the  decolorizing  action  of  acids  as  the 


PATHOGENIC    BACTERIA.  97 

tubercle  bacilli,  but  they  are  easily  stained,  requiring  but  a 
few  minutes  with  the  ordinary  watery  solutions.  They  take 
Gram's  stain  readily. 

Pathogenesis. — Arning  has  inoculated  prisoners  with  tissue 
obtained  from  leprous  patients,  and  produced  true  leprosy. 

Rabbits  which  had  been  infected  through  the  anterior  chamber 
of  the  eye  showed  the  lepra  nodules  (containing  the  lepra 
bacilli)  diffused  through  various  organs. 

In  man  the  skin  and  peripheral  nerves  are  principally  affected, 
but  the  lymphatic  glands,  liver,  and  spleen  can  also  become  the 
seat  of  the  lepra  nodules.  The  lepra  cells  which  compose  these 
nodules  contain  the  bacilli  in  large  numbers. 

Method  of  Infection. — Not  yet  determined  ;  the  air,  soil,  water, 
and  food  of  leprous  districts  have  been  carefully  examined  with- 
out result. 

Syphilis  Bacillus  of  Lustgarten  (Smegma  Bacillus  of  Alvarey 
and  Tavel).  Lustgarten  in  1885,  through  a  certain  staining 
process,  found  peculiar  bacilli  in  syphilitic  tissues  which  he 
thought  had  a  direct  connection  with  the  disease. 

In  the  same  year  Alvarey  and  Tavel  and  Matterstock  found 
a  similar  bacillus  reacting  in  the  same  way  to  Lustgarten's  color 
method  in  normal  secretions,  especially  in  the  smegrna  of  the 
prepuce. 

The  question  yet  remains  an  open  one,  what  relation  the 
syphilis  or  the  smegma  bacillus  bears  to  syphilis,  and  will 
remain  so  until  the  bacillus  can  be  cultivated,  which  so  far  has 
not  been  accomplished. 

Origin. — In  the  cells  of  syphilitic  tissue,  in  the  secretion  of 
syphilitic  ulcers,  and  in  the  smegma  of  the  prepuce  and  vulva. 

Form. — Small  slender  rods  similar  in  appearance  to  tubercle 
bacilli,  sometimes  swelled  at  the  ends  and  curved  S-shaped. 

Colorless  oval  spaces  also  present,  which  Lustgarten  calls 
spores. 

Growth. — As  before  mentioned,  they  have  not  yet  been  culti- 
vated. 

Staining. — Lustgarten's  method  : — 

1.  Aniline  water  gentian  violet,  12  to  24  hours,  and  then  2 
hours  longer  in  brood  oven. 


98  ESSENTIALS    OF    B ACTEK IOLOG Y. 

2.  Rinsed  in  alcohol.     2  to  3  minutes. 

3.  Aqueous   solution   potass,   permang.   (1£  per  cent.).     10 
seconds. 

4.  Aqueous  solution  of  sulphuric  acid.     2  seconds. 

5.  Aq.  destil.  to  wash. 

Numbers  three,  four,  and  five  repeated,  until  the  section  is 
colorless.  Then  alcohol,  oil  of  cloves,  and  Canada  balsam  as 
usual. 

De  GiacornVs  method:— 

1.  Aniline  water  fuchsia.     24  hours. 

2.  Rinse  in  dilute  tr.  ferri  clilor.  sol. 

3.  Decolorize  in  concentrated  tr.  ferri  chlor. 

4.  Wash  in  alcohol,  oil  of  cloves,  Canada  balsam,  etc. 

For  cover-glass  preparations,  wash  in  water  instead  of  alcohol. 

Tubercle  and  lepru  bacilli  are  colored  by  this  method  also, 
but  syphilis  bacilli  become  decolorized  if  washed  with  acids. 

Puthogenests. — No  pathological  actions  have  yet  been  definitely 
proven.  They  are  found  in  greater  quantities  the  younger  the 
infection  is. 

Bacillus  of  Glanders.    (Bacillus  Mallei,  Loffler-Shiitz.) 

Origin. — In  the  "  farcy  buds"  or  little  nodules  of  the  disease, 
by  Loffler  and  Shiitz  in  1882. 

Form.—  Small  slender  rods,  about  the  size  of  the  tubercle 
bacillus.  The  ends  rounded.  Never  appearing  in  large  collec- 
tions, usually  singly.  Spores  are  present. 

Properties.  —  The  rods  are  very  resistant,  living  in  a  dried  state 
for  three  months  and  longer  without  any  spores  present.  They 
are  not  motile  ;  possess,  however,  great  molecular  vibration. 

Growth. — The  growth  occurs  between  25°  and  40°  C.,  best 
at  37°  C.;  it  is  very  sparse  upon  gelatine,  but  on  glycerine-agur 
or  blood  serum  a  very  abundant  growth  occurs. 

Colonies. — On  agar  or  glycerine-agar  there  appear  in  two  to 
three  days  small  white  glistening  drops,  which  under  microscope 
seem  as  round  granular  masses  with  an  even  periphery. 

Stroke  Cultures. — On  glycerine-agar  and  blood  serum  small 
transparent  drops  of  whitish  or  grayish  color,  which  soon 
coalesce  to  form  a  broad  band. 


PATHOGENIC    BACTERIA.  99 

Potato. — An  amber-colored  honey-like  growth  which  gradually 
turns  red. 

Staining.  —  Since  the  bacillus  is  very  easily  decolorized,  some 
special  methods  have  been  recommended. 

Loffler^s.— (For  cover-glass  preparations.) 

1.  Alkaline  methylin  blue  (Loffler's).  a  5  minutes. 

2.  Acetic  acid  with  a  few  drops  of  tropseoliii.     1  second. 

3.  Washed  in  water. 

For  Sections. — Instead  of  tropseolin  acetic  acid,  the  following 
mixture  is  used  : — 

]£— Oxalic  acid  5  per  cent.  .  .  .  gtt.  j. 
Cone,  sulphuric,  acid.  ....  gtt.  ij. 
Aq.  destill. gij. —  M. 

The  sections  are  kept  in  this  5  seconds. 
Kuhne's  method.     Coverglaxs. 

1.  Warm  carbol-blue  2  min. 

2.  Decolorized  in  weak  sol.  of  muriatic  acid  (10  parts  to  500). 

3.  Washed  in  water. 
Sections  of  Tissue. 

1.  Carbol-blue,  ^  hour. 

2.  Decolorized  in  £  per  cent,  muriatic  acid. 

3.  Washed  in  distilled  water. 

4.  Dehydrated  in  alcohol  1  second. 

5.  Aniline  oil  with  6  gtts.  of  turpentine.     5  min. 

6.  Turpentine,  xylol,  Canada  balsam. 

If  contrast  stain,  add  5  gtts.  of  safranin  (Bisniark-brown)  to 
turpentine,  and  use  it  after  the  xylol. 

Pathogenesis. — If  horses,  field  mice,  or  guinea-pigs  be  inocu- 
lated subcutaneously,  with  but  a  very  small  quantity  of  culture, 
a  local  affection  results,  followed  some  time  after  by  a  general 
disturbance  ;  ulcers  form  at  the  point  of  inoculation  ;  little 
nodules,  which  then  caseate,  leaving  scars  and  involving  the 
lymphatics ;  metastatic  abscesses  then  occur  in  the  spleen  and 
lungs,  and  death  arises  from  exhaustion.  Cattle,  pigs,  and  rab- 
bits are  not  easily  affected  ;  man  is  readily  attacked. 

Manner  of  Infection.— Glanders  being  a  highly  contagious  dis- 
ease, it  requires  but  a  slight  wound  to  allow  it  to  gain  entrance. 


100        ESSENTIALS  OF  BACTERIOLOGY. 

In  horses  the  primary  sore  seems  to  be  at  the  nasal  mucous 
membrane.  In  man  it  is  usually  on  the  fingers.  Boiling  water 
or  1-10,000  sublimate  solution  will  quickly  destroy  the  virulence 
of  this  bacillus. 

Bacillus  of  Diphtheria.    (Klebs-Lbffler.) 

Origin. — In  diphtheritic  membrane,  by  Loftier,  in  1884. 

Form. — Small,  slightly  curved  rods  about  as  long  as  tubercle 
bacilli  and  twice  as  broad  ;  the  ends  are  at  times  swollen  ; 
spores  have  not  been  found. 

Properties. — They  do  not  possess  any  movement ;  do  not 
liquefy  gelatine.  They  are  not  very  resistant,  being  destroyed 
by  a  temperature  of  50°  C.,  but  they  have  lived  on  blood-serum 
five  months. 

Growth. — Grow  readily  on  all  media,  between  temperature 
of  20°  and  40°  C.  They  are  facultative  anaerobic  ;  they  grow 
quite  rapidly  and  profusely. 

Colonies  on  Gelatine  Plates.— At  24°  C.  little  round  colonies, 
under  low-power,  granular  centre  ;  irregular  borders. 

Stab  Cultures.— Small,  white  drops  along  the  needle  track.  In 
glycerine-agar  a  somewhat  profuse  growth. 

Potato.— On  alkaline  surface,  a  grayish  layer  in  48  hours. 

Blood-Serum  (after  Loffler). — Blood  serum  3  parts,  and  bouil- 
lon 1  part ;  the  bouillon  contains  peptone,  1  per  cent.  ;  chloride 
of  sodium,  £  per  cent.  ;  and  dextrin,  1  per  cent. 

On  this  medium  a  very  thick  yellowish-white  layer  occurs  on 
the  surface,  and  isolated  colonies  in  the  upper  strata. 

Staining. — Is  not  colored  by  Gram's  method.  Stained  best 
with  Loffler's  alkaline  methylin-blue. 

Pathogenesis.  —  'By  inoculation,  animals,  which  naturally  are 
not  subject  to  diphtheria,  have  had  diphtheritic  processes  de- 
velop at  the  site  of  infection  ;  hemorrhagic  oedema  then  follows, 
and  death. 

In  rabbits  paralyses  develop,  and  when  the  inoculation  occurs 
upon  the  trachea,  all  the  prominent  symptoms  of  diphtheria 
show  themselves. 

Manner  of  Infection  in  Man. — The  exact  way  is  not  yet  known. 
It  is  supposed  that  the  mucous  membrane  altered  in  some  man- 


PATHOGENIC    BACTERIA.  101 

ner,  the  diphtheria  bacillus,  then  gains  entrance  and  the  disease 
develops. 

Products. — But  it  is  not  the  mere  presence  of  the  bacillus  that 
gives  rise  to  all  trouble  ;  certain  products  which  they  generate 
get  into  the  system  and  produce  the  severe  constitutional  symp- 
toms. 

Eoux  and  Yersin,  in  1888,  discovered  that  the  injection  of  the 
filtered  culture  bouillon  (that  is,  freed  of  all  diphtheria  bacilli) 
gave  rise  to  the  same  palsies  as  when  the  bacilli  themselves  were 
introduced. 

Brieger  and  Frankel,  through  frequent  precipitation  of  the 
culture  bouillon  with  acetic  acid  and  alcohol,  obtained  a  white, 
amorphous  body,  which  gave  all  the  reactions  of  an  albumen, 
and  being  highly  toxic,  they  gave  it  the  name  of  tox-albumen. 
It  is  soluble  in  water  and  decomposed  by  higher  temperatures. 

Immunity. — Brieger  and  Frankel,  by  injecting  10  to  20  c.cm. 
of  a  three  weeks'  old  culture  of  diphtheria  bacilli,  which  had 
been  heated  at  70°  C.  for  one  hour,  produced  an  immunity  in 
guinea-pigs  against  the  virulent  form. 

Behring  found  several  ways  to  make  animals  immune.  One 
method  was  to  infect  them  with  diphtheria  and  then  inject  tri- 
chloriodine  into  them,  which  prevented  them  from  dying,  and 
they  were  then  immune. 

Site  of  Bacilli. — Bacilli  are  usually  found  in  the  older  portions 
of  the  pseudo-membrane  very  near  to  the  surface.  The  secre- 
tions of  the  throat  of  a  diphtheritic  child  produced  bacilli  three 
weeks  after  the  temperature  was  down  to  normal. 

Streptococcus  in  Diphtheria.  Streptococci  have  been  found 
quite  constant  in  diphtheria,  but  they  resemble  the  strepto- 
coccus pyogenes,  and  have  no  specific  action. 

Bacillus  of  Typhoid  or  Enteric  Fever.    (E  berth-Gaff ky.) 

Origin. — Eberth  found  this  bacillus  in  the  spleen  and  lym- 
phatic glands  in  the  year  1880,  and  Gaff  ky  isolated  and  cultivated 
the  same  four  years  later. 

Form. — Rods  with  rounded  ends  about  three  times  as  long  as 
they  are  broad.  Usually  solitary  in  tissue-sections,  but  in  arti- 
ficial cultures  found  in  long  threads.  Flagella  on  the  side. 

Properties. — They  are  very  motile  ;  they  take  the  aniline  dyes 


1C2       ESSENTIALS  OF  BACTERIOLOGY. 

less  deeply  than  some  similar  bacilli.  Spores  have  not  yet  been 
found  ;  small  oval  spaces  appear  in  some  of  the  degenerated 
bacilli  just  at  one  end,  but  these  bacilli  are  less  resistant  than 
those  without  this  so-called  spore  ;  they  do  not  liquefy  gelatine. 

FIG.  50.  FIG.  51. 


" 


,  i/  /•' 


,  Typhoid  fever  bacillus  in  pure  cul-  Bacillus  of  typhoid  fever. 

ture.    650  diameters. 

Growth.—  They  are  facultative  anaerobic  ;  grow  best  at  37° 
C.,  but  can  also  develop  at  ordinary  room  temperature.  All 
nutrient  media  can  be  used  as  culture  ground.  They  develop 
chiefly  on  the  surface,  and  very  slowly. 

Colonies  on  Gelatine  Plates.—  Two  forms  ;  the  ones  near  the 
surface  spread  out  like  a  leaf,  transparent  with  bluish  fluor- 
escence. The  deeper  ones  appear  as  whetstone  crystals  of 
uric  acid,  the  same  yellowish  tinge. 

In  five  days  they  attain  to  3  millimetres  in  diameter. 

On  Potato  Gelatine.  —  The  colonies  do  not  have  the  yellow 
color,  they  are  transparent,  later  on  they  become  dark  brown 
with  green  iridescence. 

Stab  Cultures.  —  Mainly  on  the  surface  a  pearly  layer. 

Stroke  Cultures.  —  A  transparent  thick  layer. 

Potato.  —  The  growth  here  is  quite  characteristic.  At  37°  C. 
in  48  hours  a  moist  transparent  film  is  formed  over  the  whole 
surface,  but  so  transparent  that  it  can  hardly  be  seen  without 
close  observation.  If  a  small  portion  of  this  is  placed  under  a 
microscope,  it  will  be  seen  swarming  with  bacilli. 

The  growth  never  becomes  more  prominent  ;  the  potato  must 
have  a  neutral  or  acid  reaction. 


PATHOGENIC    BACTERIA.  103 

Milk. — They  grow  very  well  in  milk  without  producing  any 
visible  changes  in  its  composition. 

Carbolized-Gelatine.— Gelatine  which  has  added  to  it  j^  per 
cent,  carbolic  acid  will  allow  the  typhoid  bacillus  to  develop, 
other  similar  bacilli  being  destroyed. 

Staining. — Colored  with  the  ordinary  aniline  dyes,  when  they 
are  warmed ;  since  they  are  easily  decolorized,  acids  should  be 
avoided. 

Grain's  method  is  not  applicable.  Tissue  sections  stained  as 
follows  : — 

Alkaline  blue 1  hour. 

Alcohol 5  seconds. 

Aniline  oil 5  minutes. 

Turpentine  oil 1  minute. 

Xylol  and  Canada  bals. 

Such  a  specimen  should  first  be  examined  with  low  power,  to 
focus  little  colored  masses,  then  examined  with  immersion  lens  ; 
these  masses  will  be  found  composed  of  bacilli. 

Similar  Bacteria.  The  Neapolitanus  bacillus  of  Emmerich  or 
faces  bacillus  of  Brieger  resembles  the  typhoid  bacillus  in  many 
ways,  the  colonies  being  the  same  and  its  structure  similar. 
But  the  growth  on  potato  is  very  different ;  a  thick,  }7ellow, 
pasty  layer  is  formed  thereon. 

In  Water.  Bacilli  have  been  found  which  also  resemble 
typhoid  bacilli,  and  one  must  be  very  careful  to  make  any 
positive  statement. 

Examination  of  Water  for  Typhoid  Bacilli. — When  a  water  is 
supposed  to  contain  typhoid  bacilli,  500  c.cm.  of  the  same  is 
mixed  with  20  gtts.  of  ^-per  cent,  carbolic  acid,  which  destroys 
many  of  the  saprophytes. 

Plates  are  then  made  as  described  under  Water  Analysis. 

Those  colonies  which  then  form  and  have  a  tendency  to  liquefy, 
are  touched  on  second  day  with  permanganate  of  potassium, 
and  when  so  colored,  destroyed  with  bichloride  of  mercury. 

Those  that  now  develop  are  transferred  by  inoculation  to  fresh 
plates.  At  the  end  of  eight  days  they  are  examined  under 
microscope  ;  every  colony  not  possessing  motile  bacilli  is  dis- 
carded. The  motile  bacilli  are  tested  with  Gram's  method  of 


104       ESSENTIALS  OF  BACTERIOLOGY. 

staining ;  those  that  do  not  take  the  stain  are  alone  retained. 
Cultures  are  made  from  these  upon  potatoes,  and,  if  the  char- 
acteristic growth  occurs,  then  only  can  they  be  called  typhoid 
bacilli  with  any  certainty. 

Patfiogenesis. — Lower  animals  have  not  yet  been  given  enteric 
fever,  though  their  death  has  been  caused  by  injection  of  the 
bacilli  into  the  veins  of  the  ear. 

In  man  it  has  been  found  in  the  urine,  blood,  sputum,  milk, 
intestinal  discharges,  roseolar  spots,  and  in  various  organs,  as 
spleen,  liver,  lymphatic  glands,  and  intestinal  villi. 

It  is  found  in  secretions  several  days  after  the  attack  has  sub- 
sided. It  is  found  only  in  this  disease,  and  regularly. 

Way  of  Infection.— The  bacilli  in  the  dejecta  of  the  diseased 
person  find  their  way  into  drinking  water,  milk,  or  dirty  clothes, 
and  so  into  the  alimentary  tract  of  a  person  predisposed  to  the 
disease.  They  enter  the  blood  through  the  lymphatics,  and  so 
become  lodged  in  various  organs. 

Products. — Brieger  found  a  ptomaine  in  the  cultures  which  he 
named  typhotoxin  with  the  formula  C9H17NO2.  It  has  no 
specific  action.  A  toxalbumen  insoluble  in  water  has  also  been 
isolated,  but,  as  experiment  animals  are  immune  to  the  disease, 
no  definite  actions  have  yet  been  determined. 

The  cultures, when  old, show  an  acid  reaction. 

Bacillus  Neapolitanus.    (Emmerich.) 

Origin. — During  the  cholera  epidemic  in  Naples,  in  1884, 
Emmerich  found  this  bacillus  in  the  blood  and  intestinal  dis- 
charges of  cholera-suffering  patients.  He  supposed  it  to  be  the 
real  cause  of  cholera  ;  but  since  then  it  has  been  shown  to  he 
nothing  more  than  the  Faeces  bacillus  which  Brieger  described, 
and  which  is  found  in  faeces  of  healthy  persons,  in  the  air  and 
various  putrefactive  processes. 

Form.— Very  much  like  the  typhoid  bacillus,  short  rods  with 
rounded  ends  with  oval  spaces  in  them  as  the  typhoid. 

Properties. — Immobile,  differing  thus  markedly  from  typhoid. 
Do  not  liquefy  gelatine. 

Growth. — They  are  facultative  anaerobic  ;  they  grow  more 
rapidly  than  the  typhoid,  and  endure  cold  and  heat  better  than 
they  do. 


PATHOGENIC    BACTERIA. 


105 


Colonies. — They  are  exactly  the  same  as  typhoid — the  same 
whetstone-shaped  deep  ones  and  the  leaf-shaped  surface  ones. 

Potato. — A  thick  yellow-brown  pasty  layer  is  formed  instead 
of  the  transparent  almost  invisible  growth  of  the  typhoid 
bacil'us. 

Staining. — Do  not  take  Gram.     Fuchsin  stains  them  easily. 

Pathogenesis. — When  large  quantities  injected  into  guinea- 
pigs,  they  die  at  times,  sometimes  with  intestinal  symptoms, 
sometimes  without. 


CHAPTER  III. 


FIG.  52. 


PATHOGENIC   BACTERIA — CONTINUED. 

Spirillum  Cholerae.    (Koch. )     Comma  bacillus  of  cholera. 

Origin. — Koch,  as  a  member  of  the  German  expedition  sent 
to  India,  in  1883,  to  study  cholera,  found  this  micro-organism 
in   the  intestinal  contents  of  cholera 
patients,  and  by  further  experiments 
identified  it  with  the  disease. 

Form.— The  microbe  as  seen  ordi- 
narily appears  as  a  short,  arc-like  body, 
about  half  the  size  of  a  tubercle  bacillus, 
but  when  seen  in  large  groups,  spirals 
are  formed,  each  little  arc  appearing 
then  as  but  a  segment,  a  vibrio;  each 
arc.  is  about  three  times  as  long  as  it 
is  broad,  and  possesses  at  each  end  a 
flagella. 

Properties. — They   are   very  motile  ; 

liquefy  gelatine.  They  are  easily  affected  by  heat  and  dryness. 
Spores  have  not  been  found,  though  some  (Hiippe)  claim  arthro- 
spores. 


Comma  bacillus,  pure  cul- 
ture.   600  diameters. 


106 


ESSENTIALS    OF    B  A  CTERIOLOG!  Y. 


FIG.  53. 


Growth. — Develops  at  ordinary  temperatures  on  all  nutrient 
media  that  have  an  alkaline  or  neutral  reaction.  They  are 
facultative  anaerobic. 

Colonies,  gelatine. — After  24  hours,  small  white  points  which 
gradually  come  to  the  surface,  the  gelatine  being  slowly  .lique- 
fied, a  funnel-shaped  cavity  formed  holding  the  colony  in  its 
narrow  part,  at  the  bottom,  and  on  the  fifth  day  all  the  gelatine 
is  liquid.  If  the  colonies  of  three  days'  growth  are  placed  under 
microscope  they  appear  as  if  composed  of  small  bits  of  frosted 
glass  with  sharp  irregular  points. 

Stab  Culture. — After  30  hours  a  growth  can  be  distinguished 
along  the  needle  track,  and  on  the  surface  a  little  cavity  has 
been  formed,  filled  up  by  a  bubble  of  air,  and  this  liquefaction 
proceeds  until  on  the  sixth  day  it  has  reached  the  sides  of  the 
tube,  tapering,  funnel-shaped  to  the  bottom  of  the  tube.  After 

several  weeks  the  spirilla 
are  found  in  little  collec- 
tions at  the  bottom  of  the 
fluid  gelatine.  In  eight 
weeks  the  bacilli  have 
perished. 

Agar. — Stroke  cultures. 
A  shiny  white  layer  lasts 
many  months. 

Potato. — A  yellow  honey- 
like  transparent  layer,  if 
the  potato  is  kept  at  ani- 
mal heat. 

Bouillon.  —  A  wrinkled 
scum  is  soon  formed  in 
bouillon.  They  live  well 
and  grow  in  sterilized  milk 
and  sterilized  water,  re- 
maining virulent  in  the 
latter  for  many  months. 

In  ordinary  water,  the  bacteria  present  are  destructive  to  the 
comma  bacillus,  and  they  die  in  a  few  days. 

Staining. — They  are  colored  well  with  watery  aniline  solu- 


Cholera  dejections  upon  a  damp  sheet,     a.- 
Formed  bacilli.    600  diameters.    (Koch.) 


PATHOGENIC    BACTERIA.  107 

tions.     The  flagella  can  be  well  seen  by  staining  according  to  the 
flagella  stain. 

Pathoyenesis. — Experiment  animals  are  not  subject  to  cholera 
Asiatica,  but  by  overcoming  two  obstacles  Koch  has  produced 
choleraic  symptoms  in  guinea-pigs.  Nicati  and  Rietsch  pre- 
vented peristalsis  and  avoided  the  acidity  of  the  stomach  juices 
by  direct  injection  into  the  duodenum,  after  tying  the  gall-duct. 
Koch  alkalinizes  the  gastric  juice  with  5  c.cm.  of  5  per  cent, 
sol.  of  sodii  carbonas,  and  then  injecting  2  grams  of  opium  tinc- 
ture for  every  300  grams  of  weight  into  the  peritoneal  cavity 
paralyzes  peristalsis.  The  cholera  culture  then  introduced 
through  a  stomach-tube,  the  animals  die  in  forty-eight  hours, 
presenting  the  same  symptoms  in  the  appearance  of  the  intes- 
tines as  in  cholera  patients,  the  serous  effusion  containing  great 
numbers  of  spirilla. 


Comma  bacillus  in  mucus,  from  a  case  of  Asiatic  cholera. 

Manner  of  Infection  in  Man. — Usually  through  the  alimen- 
tary tract,  with  the  food  or  drink,  the  intestinal  discharges  of 
cholera  patients  having  found  entrance  into  the  source  of  drink- 


108        ESSENTIALS  OF  BACTERIOLOGY. 

ing  water.  Soiled  clothes  to  fingers,  fingers  to  the  mouth,  etc. ; 
torpid  catarrhal  affection  of  the  digestive  tract  predisposing. 
The  microbe  is  not  found  in  the  blood  or  any  organ  other  than 
the  intestines,  the  tissue  of  the  small  intestines.  It  is  also 
found  in  the  vomit  and  the  intestinal  contents. 

Products. — "  Cholera  red."  When  chemically  pure  nitric  or 
sulphuric  acid  is  added  to  nutrient  peptone  cultures  of  the 
cholera  bacillus  a  rose-red  color  is  produced.  This  will  riot  take 
place  with  other  bacilli  unless  nitrous  acid  is  present.  The  cholera 
bacillus  forms  nitrites  from  the  nitrates  present  in  the  media, 
and  also  indol.  The  mineral  acid  splits  the  nitrites,  setting  free 
nitrous  acid,  which,  with  the  indol,  forms  the  red  reaction. 
This  pigment  has  been  isolated  and  extracted  and  called 
"  cholera  red."  A  ptomaine,  identical  with  cadaverin,  and  sev- 
eral other  alkaloids  have  been  obtained  from  the  cultures.  A 
toxalbumen  and  a  toxicpeptone  have  lately  been  isolated,  but 
no  special  actions  ascribed  to  them. 

Bacteria  Similar  to  the  Spirillum  of  Cholera. 

Finkler-Prior  Vibrio,  or  Spirillum  Finkleri. 

Origin.—  Found  in  the  intestinal  contents  of  a  patient  suffer- 
ing from  cholera  Asiatica  in  1884,  by  Finkler  and  Prior,  who 

thought  it  identical  with  the  spi- 
FlG-  55'  ^  rillum  of  cholera  ;  it  differs  from 

£&  it,  however,  in  many  ways,  and 
^  >vS'"/5*  nas  been  found  in  healthy  per- 

£ '  * 

Jfife  Form.— Somewhat  thicker  than 

^';;'oV  the     cholera    vibrio,     otherwise 

VMr"'  about  the  same  form  ;  it  forms 

^'  the  long  spirilla  less  often.     Has 

Flagella. 

Spirillum  Finkleri.     700  diameters.  _.     .  ... 

(FiQgge.)  Properties.—  It  is  very  motile. 

Liquefies  gelatine  in  a  short  time. 

Groicth. — It  grows  quickly  at  ordinary  room  temperature.  It 
is  facultative  aerobic. 

Colonies  o?i  Gelatine  PJates.— Round,  finely  granular  colonies, 
which  in  twenty-four  hours  are  ten  times  as  large  as  the  cholera 
colonies,  and  in  forty-eight  hours  the  whole  plate  is  liquefied, 


PATHOGENIC    BACTERIA. 


109 


FIG.  56. 


it  being  then  impossible  to  distinguish  any  separate  colonies. 
The  microscopic  appearances  in  no  way  resemble  the  cholera 
colony. 

Stab  Cultures. — The  gelatine  is  liquefied 
from  above  downwards,  like  a  stocking  in 
appearance,  and  in  three  days  completely 
liquid. 

Potato.— At  ordinary  temperature  a 
thick  gray  layer  covering  the  whole  sur- 
face. 

Water. — It  soon  perishes  in  water. 

Staining.  —  Ordinary  aniline  dyes. 

Patlicgenesis.  —  Foic  man  it  has  no  spe- 
cific action.  If  it  is  injected  into  Guinea 
pigs,  prepared  as  described  under  the 
cholera  bacillus,  they  die,  the  intestines 
having  a  foul  odor,  and  the  bacilli  then 
found  in  great  numbers. 

Spirillum  Tyrogenum.    (Den eke.) 

Origin. — In  1885  Deneke  found  in  old 
cheese  a  spirillum  very  similar  in  appear- 
ance to  the  cholera  spirillum. 

Form. — The  same  as  the  cholera  vibrio. 

Properties. — Very  motile,  liquefy  gela- 
tine. 

Growth. — They  grow  quicker  than  the 
cholera,  and  slower  than  the  Finklcr  ;  they  are  also  facultative 
aerobic. 

Colonies.— They  at  first  resemble  cholera  colonies  ;  they  have, 
however,  a  yellow-green  iridescence,  and  are  somewhat  more 
irregular  ;  also  grow  more  rapidly. 

Slab  Cultures.— A  thick  line  along  the  needle-track  and  the 
yellow  colonies  forming  at  the  bottom,  on  the  surface  a  bubble 
of  air  similar  to  the  cholera.  The  gelatine  is  all  liquid  in  two 
weeks. 

Potato. — At  brood-heat  a  thin  yellow  membrane,  but  not 
always  constant. 

Staining,  as  cholera  bacillus. 


Stab  Culture.     (Finklcr- 
Prior.) 


110  ESSENTIALS    OF    BACTERIOLOGY. 

PathcHjenesis.—  When  injected  into  animals  prepared  as  for  the 
cholera  bacillus,  a  certain  number  die. 

Vibrio  Metschnikovi.  (Gamaleia.) 

Origin. — In  the  intestines  of  fowls  suffering 'from  a  gastro- 
enteritis, common  in  Kussia.  Gamaleia  found  a  spirillum  which 
bears  so  close  a  resemblance  to  the  cholera  bacillus,  both  in  form 
and  growth,  that  it  cannot  be  distinguished  by  these  character- 
istics alone. 

Form. — As  cholera  bacillus. 

Growth. — Two  kinds  are  found  on  the  gelatine  plate — one  that 
is  identical  in  appearance  with  the  cholera  colony,  the  other  more 
liquefying,  resembling  the  Finkler  spirillum.  If  now  a  second 
plate  be  inoculated  from  either  one  of  these  forms,  both  kinds 
again  are  found  grown,  so  that  it  is  not  a  mixture  of  two  bacilli. 

Stab  Culture.— Similar  to  the  cholera  growth,  a  trifle  faster  in 
growing. 

Staining. — As  cholera. 

Pathogenesis. — To  differentiate  it  from  cholera,  these  bacilli, 
when  injected  into  animals,  prove  very  fatal,  and  no  especial 
precautions  need  be  taken  to  make  the  animal  susceptible.  In 
the  pigeon,  guinea-pig,  and  chicken  it  produces  a  hemorrhagic 
oedema,  and  a  septicaemia  which  has  been  called  "  Vibrion 
septicccmia."  The  blood  and  organs  contain  the  spirilla  in 
great  numbers. 

Products. — The  nitrites  are  formed  just  as  in  cholera  bacillus, 
and  the  red  reaction  given  when  mineral  acids  added  to  gelatine 
cultures.  Certain  products  also  which,  when  injected,  give 
immunity.  The  cultures  are  first  heated  for  one  half  hour  at 
100°  C.,  which  destroys  the  germs,  and  then  this  sterilized  pro- 
duct injected.  (5  c.cm.  of  a  five  days'  old  sterilized  culture.) 

In  a  couple  of  weeks  1  to  2  c.cm.  of  the  infected  blood  can  be 
injected  without  causing  any  fatal  result. 

Bacteria  of  Pneumonia.  Two  forms  of  bacteria  have  been 
found  in  this  disease,  and  thought  at  different  times  to  be  the 
cause  of  the  same. 

Neither  one  of  them  is  constant  in  pneumonia  ;  and  since 
many  other  pathological  processes  have  shown  them  they  can 
hardly  be  set  down  as  the  sole  cause  of  pneumonia. 


PATHOGENIC    BACTERIA.  Ill 

Klebs  in  1875  called  attention  to  the  presence  of  bacteria  in 
pneumonia,  and  in  1882  Friedlander  developed  a  bacillus  from 
the  lung  tissue  of  a  pneumonic  person,  which  he  thought  was  a 
coccus,  and  called  it  pneumococcus. 

In  1886  A.  Frankel  and  Weichselbaum  proved  that  this 
microbe  was  not  constant,  in  fact  was  rare. 

A.  Frankel  obtained  in  the  majority  of  cases  of  pneumonia  a 
microbe  that  he  had  described  in  1884  under  the  name  of 
sputum-septicaemia  micrococcus. 

Weichselbaum  now  called  it  "  Diplococcus  Pneumonia,"  and 
believed  it  to  be  the  real  cause  of  pneumonia.  It  has  been  found 
in  many  other  serous  inflammations,  and  also  in  the  mouth  of 
healthy  persons. 

Streptococcus  pyogenes  and  staphylococcus  pyogenes  aureus  have 
been  found  in  some  cases. 

FIG.  57. 


Pneumo-bacillus  of  Friedlander,  with  capsule. 

Pneumo-bacillus  (Pneumococeua).     (Friedlander.) 

Origin. — In  the  lung  of  a  croupous-pncumonia  person,  by 
Friedlander,  in  1882. 

Form.— Small,  almost  oval-shaped  rods,  nearly  as  wide  as 
they  are  long  ;  often  in  pairs,  they  were  at  first  believed  to  be 
cocci.  In  bouillon  cultures  the  rod-form  becomes  more  visible. 
In  tissues  each  bacillus  is  surrounded  by  a  faint  capsule  ;  but 
not  around  those  developed  in  artificial  cultures.  Spores  have 
not  been  found. 

Properties. — They  are  immobile  ;  do  not  liquefy  gelatine.  A 
gas  is  produced  in  gelatine  cultures. 

Growth.  —  Grows  rapidly  on  all  media  at  ordinary  temperature  ; 
is  facultative  aerobic. 

Colonies.— On  gelatine  plates.  Small  white  round  colonies, 
reaching  the  surface  in  the  course  of  three  or  four  days  ;  appear- 


112 


ESSENTIALS  OF  BACTERIOLOGY. 


Fro.  58. 


ing  then  as  little  buttons,  with  a  porcelain-like  shimmer,  the 
edges  smooth. 

Stab  Culture.— A  growth  along  the  needle-track,  but  on  the 
surface  a  button-like  projection,  which  gives  to  the  growth  the 
appearance  of  a  nail  driven  into  the  gelatine, 
its  head  resting  on  the  surface  ;  therefore 
such  cultures  are  called  "  Nail  cultures." 
See  Fig.  58.  Old  cultures  are  colored  brown, 
and  contain  bubbles  of  gas. 

Potato.  — A  yellow,   moist  layer  in  a  few 
days  at  brood-heat.     Gas  bubbles  develop. 

Staining.— The    ordinary    aniline    stains. 
The  sections  do  not  take  Gram's  method  ; 
are  therefore  not  suited  for  double  staining. 
Capsule. — Stained  as  follows  : — 
Cover  glasses. 

1.  Acetic  acid,  two  minutes. 

2.  Allow  acetic  acid  to  dry  by  blowing  air 
upon  it  through  a  glass  tube. 

3    Saturated,  aniline  water.    Gent,  violet, 
ten  seconds. 

4.  Rinse  in  water.  Mount  in  Canada  balsam. 
For  Sections. 


Bacillus  of  Pneumo- 
nia. Stab  Culture. 
(Nail  Culture.) 


cone.  ale.  gent,  violet,  50.0 


1.  Stain  in  warm  ^  aqua,  100.0 

acetic  acid,  10. 

for  24  hours.  M. 

2.  Rinse  in  one  per  cent,  acetic  acid. 

3.  Alcohol  to  dehydrate.    Mount  in  balsam. 

The  capsule  will  be  found  stained  a  light  bine,  the  bacillus  a 
deep  blue. 

Pathogentsis.— Animals  are  not  affected  unless  the  culture  is 
injected  intrapleura. 

Pneumobacilhis  of  Frankel.   (A.  Frankel  and  Weichselbaum.) 
Synonyms. — Pneumococcus  ;  Diplococcus  of  Pneumonia  ;  Mi- 


PATHOGENIC    BACTERIA. 


113 


crococcus  of  sputum  septicaemia  ;  Micrococcus  Pasteuri  ;  Diplo- 
coccus  lanceolatus. 

Origin. — A,  Fraukel  found  it  in  the  sputum  of  pneumonic 
patients,  thinking  it  at  first  to  be  the  micrococcus  of  sputum 
septicaemia  ;  later  he  believed  it  to  be  the  cause  of  pneumonia. 

Form. — Oval  cocci  they  were  at  first  called,  but  they  are  now 
known  to  be  rod-shaped,  being  somewhat  longer  than  broad  ; 
varying,  however,  much  in  size  and  shape.  Usually  found  in 
pairs,  sometimes  in  filaments  of  three  and  four  elements.  In 
the  material  from  the  body  a  capsule  surrounds  each  rod.  In 
the  artificial  cultures  this  is  not  found. 


FIG.  59. 


Bacillus  of  Pneumonia  in  Saliva.    (After  Biondl.) 

Properties.  —They  are  without  self-movement ;  do  not  liquefy 
gelatine. 

Growth.— Grow  only  at  high  temperature,  35°  C.  ;  are  facul- 
tative anserobic.  The  culture  media  must  be  slightly  alkaline  ; 
the  growth  is  slow. 

Colonies  on  Gelatine  Plates. — Since  the  temperature  must  be 
somewhat  elevated,  the  gelatine  media  need  to  be  thicker  than 
usual  (15  per  cent,  gelatine),  in  order  to  keep  it  solid,  and  a 
temperature  of  24°  C.  used.  Little  round  white  colonies,  some- 
what granular  in  the  centre,  growing  very  slowly. 
8 


114  ESSENTIALS    OF    BACTERIOLOGY. 

Stab  Cultures. — Along  the  needle-track  small  separate  white 
granules,  one  above  the  other,  like  a  string  of  beads. 

Stroke  Culture.  — On  agar,  transparent,  almost  invisible  little 
drops  resembling  dew  moisture. 

Bouillon. — They  grow  better  here  than  in  the  other  media, 
remaining  alive  a  longer  period  of  time. 

Staining. — Takes  Gram's  method  and  the  other  aniline  stains 
very  readily.  The  capsule  stained  the  same  way  as  that  of  the 
Friedlande-r  bacillus. 

Pathogenesis. — Rabbits  and  guinea-pigs,  if  subcutaneously  in- 
jected, die  in  the  course  of  a  couple  of  days  with  septicaemia. 
(0.1  c.cm.  of  a  fresh  bouillon  culture  suffices.) 

Autopsy  shows  greatly  enlarged  spleen  and  myriads  of  bacilli 
in  the  blood  and  viscera,  the  lungs  not  especially  affected.  If 
injected  per  trachea,  a  pneumonia  occurs.  In  man  in  90  pel- 
cent,  of  croupous  pneumonia  they  are  found  and  usually  only 
during  the  existence  of  the  u  prime  juice"  sputum,  i.  e.,  the  first 
stage. 

Fia.  60. 


/  ,x 


* 
t 


Micrococcus  tetragenus  in  sputum  (tubercle  bacillus  also). 

They  have  also  been  found  in  pleuritis,  peritonitis,  pericarditis, 
meningitis,  and  endocarditis.  They  stand  in  some  intimate  re- 
lation with  all  infectious  inflammations  of  the  body.  Their 
presence  in  healthy  mouth  secretion  does  not  speak  against  it,  it 


PATHOGENIC    BACTERIA. 


115 


Fio.  61. 


requiring  some  slight  injury  to  allow  this  ever-present  germ  to 
develop  the  disease. 

Bacillus  of  Rhinoscleroma,  (Frisch.  1882.)  It  was  found  in 
the  tissue  of  a  rhinoscleroma,  but  resembles  the  Friedlander 
bacillus  in  nearly  every  respect,  and  as  the  disease  rhinoscleroma 
was  not  reproduced  by  the  inoculation  of  the  bacillus  in  animals, 
it  can  be  considered  identical.  The  growth,  cultures,  and  pro- 
perties are  the  same  as  the  pneumobacillus  of  Friedlauder. 

Micrococcus  Tetrageims.    (Koch.    Gaffky). 

Origin. — Koch  found  this  microbe  in  the  cavity  of  a  tubercu- 
lous lung.     Gaft'ky,  in  1883,  studied  its  patho- 
genic actions  and  gave  it  the  name  it  now 
bears. 

.Form. — Cocci  which  are  gathered  in  the  tis- 
sues in  groups  of  four,  forming  a  square,  a 
tetrad.  See  Fig.  62.  In  artificial  culture, 
sometimes  found  in  pairs.  A  capsule  of  light 
gelatinous  consistence  surrounds  each  tetrad. 

Properties.— They  are  immobile  ;  do  not 
liquefy  gelatine. 

Growth. — They  grow  well  on  all  nutrient 
media  at  ordinary  and  brood  temperatures ; 
are  facultative  aerobic.  They  grow  slowly. 

Colonies  in  gelatine  plates.  In  two  days, 
little  white  spots,  which  when  on  the  surface 
form  little  elevations  of  a  porcelain-like  ap- 
pearance ;  under  low  power  they  are  seen  very 
finely  granulated. 

Stab  Culture.— Small  round  separated  colo- 
nies along  the  needle-track,  and  on  the  sur- 
face a  button-like  elevation,  a  form  of  "nail 
culture."  See  Fig.  58. 

Potato. — A  thick  slimy  layer  which  can  be 
loosened  in  long  shreds. 

Staining. — Colored  with  the  ordinary  aniline 
stains.  Gram's  method  also  applicable. 

Pathogenesis. — White  mice  and  guinea-pigs         stab  Culture, 
die  in  a  few  days  of  septicaemia  when  injected      Micro^8  tetra~ 


116       ESSENTIALS  OF  BACTERIOLOGY. 

with  the  tetragenus  cultures,  and  the  mierococcus  is  then  found 
in  large  numbers  in  the  blood  and  viscera. 

Field  mice  are  immune. 

In  the  cavities  of  tubercular  lungs,  in  the  sputum  of  phthisical 
and  healthy  patients,  it  is  often  found,  but  what  action  it  has 
upon  man  has  not  yet  been  determined. 

Capsule  Bacillus.    Pfeitfer. 

Origin.  —Stringy  exudate  and  blood  of  a  dead  guinea-pig. 

Form. — Thick  little  rods,  sometimes  in  long  threads.  Large 
oval  capsules  in  the  stained  preparations. 

Properties. — Immotile,  not  liquefying,  an  odorless  gas  in  gela- 
tine cultures. 

Growth. — At  ordinary  temperatures,  quite  rapidly  ;  facultative 
anserobin. 

Gelatine  Plates. — Oval  points,  and  like  a  porcelain  button  on 
the  surface. 

Stub  Cultures. — Like  the  pneumonia  bacillus  of  Friedlander. 

Potatoes.— Abundant  growth,  yellow  color  and  moist,  coming 
off  in  strings. 

Staining. — Hot  fuchsin  colors  the  capsule  intensely  ;  then  care- 
fully decolorizing  with  acetic  acid,  the  capsules  are  seen  rad  or 
light  violet  around  the  deeply-tinged  bacillus.  Gram's  method 
not  applicable. 

Patlwgenesis. — Subcutaneously  injected  in  mice,  they  die  in 
48  hours.  Rabbits  die  whon  a  large  quantity  is  injected  into 
the  circulation.  The  blood  and  juices  have  a  peculiar  stringy, 
fibrinous  consistence. 

Micro-Organisms  of  Suppuration.  The  suppuration  of  wounds 
is  due  to  the  presence  of  germs.  The  knowledge  of  this  fact  is 
the  basis  of  the  antiseptic  treatment  in  surgery  ;  for  when  the 
microbes  can  be  destroyed  or  their  entrance  prevented,  the 
wounds  are  made  clean  and  kept  without  suppurating.  Vari- 
ous forms  of  bacteria  have  been  found  in  septic  procer-ses,  and 
the  formation  of  pus  cannot  be  ascribed  to  any  particular  one 
alone ;  some,  more  common  than  others,  are  found  in  nearly  all 
forms  of  suppuration  ;  others  give  rise  to  special  types. 

Wounds  are  often  irritated  by  foreign  bodies  and  chemicals, 
and  a  discharge  occurs  in  them  even  when  every  aseptic  and 


PATHOGENIC    BACTERIA. 


117 


antiseptic  precaution  has  been  taken ;  but  such  a  discharge  is 
free  from  bacteria,  and  no  more  like  pus  than  a  benign  growth 
is  like  a  malignant  one. 

Streptococcus  Pyogenes.  (Bosenbach.)  Streptococcus  erysipe- 
latis.  (Fehleisen. ) 

Origin.—  Fehleisen  discovered  this  microbe  in  the  lymphatics 
of  the  skin  in  erysipelas,  and  he  thought  it  the  cause  of  the 
same.  Under  the  name  streptococcus  pyogenes,  Kosenbach 

FIG.  62. 


Streptococcus  pyogenes  in  section  of  skin. 

described  an  identical  coccus  which  has  been  found  in  nearly 
all  suppurative  conditions. 

Form. — Small  cocci  singly  and  in  chain-like  groups.  Spores 
have  not  been  found,  though  it  is  supposed  because  of  their 
permanency  that  spores  are  present. 

Properties. — They  are  immotile,  do  not  liquefy  gelatine. 

Growth. — They  grow  slowly,  usually  on  the  surface,  and  best 
at  higher  temperatures. 

Colonies. — In  three  days  a  very  small  grayish  speck,  which 
hardly  ever  becomes  much  larger  than  a  pin-head  ;  under  micro- 
scope, looking  yellowish,  finely  granular,  the  edges  quite  defined. 

Stab  Cultures. — Along  the  needle-track  little  separated  colonies 
like  strings  of  beads,  which  after  a  time  become  one  solid  white 
string. 

Stroke  Culture. — Little  drops,  never  coalescing,  having  a  bluish 
tint. 


118        ESSENTIALS  OF  BACTERIOLOGY. 

Potato.— No  apparent  growth. 

Bouillon. — At  37°  C.  clouds  are  formed  in  the  bouillon,  which 
then  sink  to  the  bottom,  and  long  chains  of  cocci  found  in  this 
growth. 

Staining. — Easily  colored  with  the  ordinary  stains.  Gram's 
method  is  also  applicable. 

Pathogenesis. — Inoculated  subcutaneously  in  the  ear  of  a 
rabbit,  an  erysipelatous  condition  develops  in  a  few  days, 
rapidly  spreading  from  point  of  infection. 

In  man,  inoculations  have  been  made  to  produce  an  eft'ect 
upon  carcinomatous  growths.  Erysipelas  was  always  produced 
thereby.  When  it  occurs  upon  the  valves  of  the  heart,  endo- 
carditis results.  Puerperal  fever  is  caused  by  the  microbe  in- 
fecting the  endometrium,  the  Streptococcus  puerperalis  of  Frankel 
being  the  same  germ. 

In  scarlatina,  variola,  yellow  fever,  cerebro-spinal  meningitis, 
and  many  similar  diseases,  the  microbe  has  been  an  almost  con- 
stant attendant. 

In  erysipelas  the  cocci  reside  in  the  lymphatic  glands  and 
ducts.  They  have  not  been  found  in  the  blood.  In  air,  soil,  and 
putrefying  matters  they  have  been  often  discovered. 

Staphylococcus  Pyogenes  Aurens.    (Boscnbach.) 

Origin. — Found  very  commonly  in  pus  (80  per  cent,  of  all  sup- 
purations), in  air,  water,  and  earth  ;  also  in  sputum  of  healthy 
persons. 

FIG.  63. 


1.  (48  hours.)         Colonies  of  micrococcus  pyogenes  aureus.         2.  (5  days.) 

Form. — Micrococci  in  clusters  like  bunched  grapes,  hence  the 
name  staphylo,  which  means  grape.  They  never  form  chains. 
Spores  have  not  been  found,  though  the  cocci  are  very  resistant. 


PATHOGENIC    BACTERIA. 


119 


FIG.  Gl. 


Properties.  —Without  movement ;  liquefying  gelatine.  It  gives 
rise  to  an  orange-yellow  pigment  in  the  various  cultures. 

Growth. — It  grows  moderately  fast  at  ordinary  temperature, 
and  can  live  without  air,  a  facultative  serobin  and  anuerobin. 

Colonies  on  Gelatine. — On  second  day  small  dots  on  the  surface, 
containing  in  their  centre  an  orange-yellow  spot.  The  gelatine 
all  around  the  colony  is  liquefied  ;  the  size  is  never  much  greater 
than  that  attained  the  second  day. 

Colonies  on  Agar. — The  pigment  remains  a 
long  time. 

Stab  Culture. — At  first,  gray  growth  along 
the  track,  which,  after  three  days,  has  settled 
at  the  bottom  of  the  tube  in  a  yellow  granular 
mass,  the  gelatine  being  all  liquid. 

Stroke  Culture  on  Agar. — The  pigment  dif- 
fused over  the  surface  where  the  growth  is,  in 
moist  masses. 

Potato. — A  thin  white  layer  which  gradu- 
ally becomes  yellow  and  gives  out  a  doughy 
smell. 

Staining. — Very  readily  colored  with  ordi- 
nary stains  ;  also  with  Gram's  method. 

Pathogenesis. — When  rabbits  are  injected 
with  cultures  of  this  microbe  into  the  knee- 
joint  or  pleura,  they  die  in  a  day.  If  injected 
subcutaneously,  only  a  local  action  occurs, 
namely,  abscesses. 

If  directly  into  circulation,  a  general  phleg- 
monous  condition  arises,  the  capillaries  become 
plugged  with  masses  of  cocci,  infarct  occur  in  kidnej^  and  liver, 
and  metastatic  abscesses  form  in  viscera  and  joints.     Garre,  by 
rubbing  the  culture  on  his  forearm,  caused  carbuncles  to  appear. 

Fracturing  a  long  bone  in  an  animal  and  then  injecting  the 
staphylococcus  into  a  large  vein,  as  the  jugular,  will  produce 
osteomyelitis.  Becker  isolated  this  microbe  from  several  cases 
of  osteomyelitis,  and  thought  it  a  specific  germ,  giving  it  the 
name  of  4'  micrococcus  of  osteomyelitis. " 

Suppuration  is  nearly  always  produced  by  this  microbe,  and 
it  is  found  in  the  majority  of  suppurative  processes. 


Stab  culture.  Micro- 
coccus  pyogenes 
aureus. 


120       ESSENTIALS  OF  BACTERIOLOGY. 

Micrococcus  Pyogenes  Albus.  (Kosenbach. )  Similar  in  every 
respect  to  the  pyogenes-aureus,  except  that  it  does  not  form  a 
pigment. 

Micrococcus  Pyogenes  Citrens.  (Passet.)  This  staphylo- 
coccus  liquefies  gelatine  less  rapidly  than  the  pyogenes  aureus, 
and  forms  a  citron-yellow  pigment  instead  of  the  orange-yellow 
of  the  aureus. 

Micrococcus  Cereus  Albus.  (Passet.)  Differs  from  the  pyo- 
genes albus  in  the  form  of  colony.  A  white  shiny  growth  like 
drops  of  wax;  hence  the  name  cereus.  It  was  found  in  pus,  but 
gave  no  action  in  animals. 

Micrococcus  Cereus  Flavus.  (Passet.)  A  lemon-yellow 
colored  growth  after  a  sbort  time,  otherwise  not  differing  from 
cereus  albus. 

Micrococcus  Pyogenes  Tenuis.    (Rosenbach.) 

Origin.—  Found  in  the  pus  of  large  inclosed  abscesses. 

Form.— Cocci,  without  any  especial  arrangement. 

Properties.— Not  much  studied. 

Growth. — It  was  cultivated  on  agar,  on  which  it  formed  in 
clear  thin  colonies ;  along  the  needle-track  an  opaque  streak, 
looking  as  if  varnished  over. 

Bacillus  Pyocyaneus.    (Gessard.) 

Synonyms. — Bacterium  a3ruginosum,  bacillus  fluorescens. 
(Schroter.)  The  bacillus  of  bluish-green  pus. 

Origin. — Found  in  1882  in  the  green  pus  in  pyocysemia. 

Form.  —  Small  slender  rods  with  rounded  ends,  easily  mistaken 
for  cocci.  Often  in  groups  of  four  and  six,  without  spores. 

Properties.— Very  motile  ;  liquefy  gelatine  rapidly  ;  a  peculiar 
sweetish  odor  is  produced  in  the  cultures,  and  a  blue  pigment. 

Growth. — Develops  readily  at  ordinary  temperature,  growing 
quickly  and  mostly  on  the  surface  ;  it  is  an-obic.  Colonies  on  gela- 
tine plate.  In  two  or  three  days  a  greenish  iridescence  appears 
over  the  whole  plate,  the  colonies  having  a  funnel-shaped  lique- 
faction, and  appearing  under  low  power  when  still  young,  as 
yellowish  green,  the  periphery  being  granulated.  - 

Stab  Cultures. — Mainly  in  upper  strata,  the  liquefaction  funnel- 
shaped,  the  growth  gradually  settling  at  the  bottom,  a  rich  green 
shimmer  forming  on  the  surface,  and  the  gelatine  having  a  deep 
fluorescence. 


PATHOGENIC    BACTERIA.  121 

Potato. — The  potato  is  soaked  with  the  pigment,  a  deep  fold 
of  green  occurring  on  the  surface. 

Staining. — With  ordinary  aniline  dyes. 

Pathogenesis. — When  animals  are  injected  with  fresh  cultures 
in  the  peritoneal  cavities  or  cellular  tissues,  a  rapidly  spreading 
oedema  with  general  suppuration  develops.  The  bacilli  are 
then  found  in  the  viscera  and  blood. 

If  a  small  quantity  is  injected,  a  local  suppuration  occurs,  and 
if  the  animal  does  not  die  it  then  can  withstand  large  quanti- 
ties. It  is  immune. 

The  Pigment.  Pyocyanin. — When  the  pus,  bandages,  and 
dressings  containing  the  bacillus  pyocyaneus  are  washed  in 
chloroform,  the  pigment  is  dissolved  and  crystallizes  from  the 
chloroform  in  long  needles.  It  is  soluble  in  acidulated  water, 
which  is  turned  red  thereby,  and  when  neutralized  the  blue  color 
returns.  It  has  no  pathogenic  action.  It  is  an  aromatic  com- 
pound. The  bacillus  has  no  especial  action  on  the  wound,  and 
is  found  sometimes  in  perspiration  of  healthy  persons. 

Bacillus  Pyocyaneus.  |3.  (Ernst.)  A  bacillus  found  in  gray- 
ish pus-colored  bandages. 

The  only  especial  difference  between  this  and  the  above  is  the 
formation  of  brownish-yellow  pigment  instead  of  pyocyanin.  The 
form  and  appearance  of  cultures  otherwise  the  same. 

Micrococcus  Gonorrhoea.     Gonococms.     (Neisser.)    In  1879 
Neisser  demonstrated  the  presence  of  this 
germ  in  the  secretion  of  specific  urethritis.  'IG'      ' 

Form.— Cocci,  somewhat  triangular  in 
form,  found  nearly  always  in  pairs,  the  base 
of  one  coccus  facing  the  base  of  the  other, 
and  giving  the  appearance  of  a  Vienna  roll, 
hence  the  German  name  Seinmel  (roll)-form. 
Four  to  twelve  such  pairs  are  often  found 
together. 

Properties. — No  movement  of  their  owrn. 
Culture. — On  gelatine-agar  or  potato  they  Gonococci  in  gon- 
do  not  grow,  and  only  upon  human-blood  orrhoeai  pus.  Ani- 
serum  have  they  given  any  semblance  of  a 
growth.  The  temperature  must  be  between 
33°  and  37°  C.,  and  the  growth  occurs  very  slowly  and  sparsely. 


122       ESSENTIALS  OF  BACTERIOLOGY. 

In  three  days  a  very  thin,  almost  invisible,  moist  yellowish 
growth,  seeming  to  be  composed  of  little  drops. 

Under  low  power  small  processes  are  seen  shooting  out  from 
the  smooth  border. 

FIG.  66. 


(Jonococci  in  pus. 

It  requires  to  be  then  transferred  to  fresh  media,  as  it  quickly 
perishes. 

Staining.— Colored  easily  with  all  ordinary  aniline  stains. 

Gram's  method  is  not  ai>plicable,  this  being  one  of  its  main 
diagnostic  features. 

The  following  method  for  coloring  cover-glasses  is  recom- 
mended by  Neisser. 

The  cover-glasses,  with  some  of  the  urethral  discharge  smeared 
upon  them,  are  covered  with  a  few  drops  of  alcoholic  solution  of 
eosin  and  heated  for  a  few  minutes  over  the  flame.  The  excess 
of  the  dye  is  removed  with  filter  paper,  then  the  cover-glass 
placed  in  concentrated  methylin  blue  (alcoholic  solution)  for  15 
seconds,  and  rinsed  in  water. 

The  gonorocci  nre  dark  blue,  the  protoplasm  of  the  cell  pink, 
and  the  nucleus  a  light  blue,  the  gonococci  lying  in  the  proto- 
plasm next  to  the  nucleus. 

Other  bacteria  are  similar  to  the  gonococci  in  form  ;  the}'  are 


PATHOGENIC    BACTERIA.  123 

distinguished  from  the  gonococcus,  in  that  they  are  colored  with 
Gram's  method,  whereas  the  micrococcus  of  gonorrhoea  is  not. 
Therefore  it  is  always  necessary,  after  having  first  found  these 
peculiar-shaped  microbes,  to  apply  Gram's  stain,  and  if  they  are 
then  not  found  one  can  safely  say  it  is  the  gonococcus. 

Pathogenesis. — The  attempts  to  infect  the  experiment  ani- 
mals with  gonorrhoea  have  so  far  been  without  success.  In  man, 
upon  a  healthy  urethra,  a  specific  urethritis  was  produced  with 
even  the  20th  generation  of  the  culture.  Gonorrhoeal  ophthalmia 
contains  the  cocci  in  great  numbers,  and  gonorrhoeal  rheumatism 
is  said  to  be  caused  by  the  lodgment  of  the  cocci  in  the  joints. 

The  microbes  have  been  found  long  after  the  acute  attack, 
when  only  a  very  slight  oozing  remained,  and  the  same  were 
very  virulent. 

The  specific  inflammations  of  the  generative  organs  of  the 
female  are  due  to  this  microbe  extending  its  influence,  having 
gained  entrance  through  the  vagina.  It  is  found  chiefly  in  the 
superficial  layers  of  the  mucous  membrane. 

Similar  Microbes  found  in  the  Urethra  and  Vagina. 

Micrococcus  Citreus  Conglomerates.  (Bumm.)  Very  similar 
to  the  gonococci  in  form,  they  are,  however,  easily  cultivated, 
and  form  yellow  colonies  which  dissolve  the  gelatine  and  grow 
quite  rapidly  \  the  surface  of  the  gelatine  is  at  first  moist  and 
shiny,  but  later  on  wrinkled.  They  are  colored  with  Gram's 
method,  and  have  no  special  pathological  action.  They  are  found 
in  the  air  and  gonorrhoeal  pus. 

Diplococcus  Albicans  Amplus.  (Bumm. )  In  vaginal  secretion. 
The  diplococci  are  much  larger  than  the  gonococci,  but  similar 
in  form.  They  are  also  cultivated  upon  gelatine  plates,  grayish- 
white  colonies,  which  slowly  liquefy  gelatine.  They  grow  mode- 
rately rapid.  Stained  with  Gram's  method,  and  have  no 
pathogenic  action. 

Diplococcus  Albicans  Tardissimus.    (Bumm.) 

Origin. — In  urethral  pus. 

Form. — Like  gonococci. 

Properties.— Im motile  ;  do  not  liquefy  gelatine. 

Growth. — Very  slow  at  ordinary  temperature,  but  more  rapid 


124  ESSENTIALS    OF    13  ACTEKIOLOG  Y  . 

at  brood-heat.     The  colonies  are  as  small  white  points,  which 
under  low  power  appear  brown  and  opaque. 

Agar  Stroke  Culture.— Grayish-white  growth,  which  after  two 
months  is  like  a  skin  upon  the  surface. 

Staining.— Takes  Gram's  method. 

Pathogenesis.—Noue  known.  . 

Micrococcus  Subflavus.    (Bumm.) 

Origin. — In  lochial  discharges,  in  vagina  and  urethra  of 
healthy  persons. 

Form. — As  gonococci. 

Properties.—  Not  motile  ;  liquefy  gelatine  slowly  ;  a  yellow- 
brownish  pigment. 

Growth.— Grows  slowly  on  all  media,  forming  on  gelatine, 
after  two  weeks,  a  moist  yellowish  surface  growth. 

Potato.  —  Small  half-moon-shaped  colonies  which,  after  three 
weeks,  become  light-brown  in  color,  and  covering  the  surface  as 
a  skin. 

Staining. — Colored  with  Gram. 

Pathogenesis. — Not  acting  upon  the  mucous  membrane,  but 
when  injected  in  cellular  connective  tissue,  an  abscess  results 
which  contains  myriads  of  diplococci. 

The  gonococcus  is  distinguished  from  all  these  similar  micro- 
cocci  by  being  found  usually  within  the  cell  protoplasm. 

Secondly.— Not  stained  with  Gram's  method. 

Thirdly. — lief  using  to  grow  readily  upon  gelatine. 

All  the  similar  bacteria  being  easily  cultivated. 

These  characteristics,  taken  in  tolo,  form  sufficient  features  for 
its  ready  recognition,  and  as  it  is  often  a  serious  question  to 
decide,  not  so  much  because  of  the  patient's  health  as  because 
of  his  character,  we  should  be  very  careful  not  to  pronounce  a 
verdict  until  we  have  tested  the  micro-organism  as  above.  AVlu  n 
the  germ  so  tested  is  found,  the  process  can  be  called  vj,<  -iji 
without  a  doubt. 

Bacillus  of  Tetanus.    (Nicolaicr-Kitasato.) 

Origin.—  Nicolaier  found  this  bacillus  in  the  pus  of  a  wounH 
in  one  who  had  died  of  tetanus,  describing  it  in  1884. 

Kitasato  has  since  then  been  able  to  isolate  and  cultivate  this 
germ.  (1889.) 


PATHOGENIC    BACTERIA. 


125 


Form. — A  very  delicate,  slender  rod.  somewhat  longer  than 
the  bacillus  of  mouse  septicaemia,  which  is  the  smallest  bacillus. 

When  the  spores  form,  a  small  swelling  occurs  at  the  end 
where  the  spore  lies,  giving  it  a  drum-stick  shape. 

FIG.  67. 


Bacillus  of  Tetanus  with  spores. 

Properties.  —  Not  very  motile,  though  distinctly  so ;  liquefies 
gelatine  slowly.  The  cultures  give  rise  to  a  foul-smelling  gas. 

Growth.—  Develops  very  slowly,  best  at  brood-heat  (36°  to  38° 
C.),  and  only  when  all  oxygen  is  excluded,  an  obligatory  ancero- 
bin.  In  an  atmosphere  of  carbon  dioxide  gas  it  cannot  grow, 
but  in  hydrogen  it  flourishes. 

Colonies  on  gelatine  plates  in  an  atmosphere  of  hydrogen. 
Small  colonies.  After  four  days  a  thick  centre  and  radiating 
wreath-like  periphery,  like  the  colonies  of  bacillus  subtilis. 

High  Stab- Culture. — (The  gelatine  having  2  per  cent,  glucose 
added  and  filling  the  tube.)  Along  the  lower  portion  of  the  needle- 
track,  a  thorny-like  growth,  little  needle-like  points  shooting 
out  from  a  straight  line.  The  whole  tube  becomes  clouded  as 


126 


ESSENTIALS    CF    li ACTERIOLOGY. 


the  gelatine  liquefies,  and  then  the  growth  settles  at  the  bottom 
of  the  tube. 

FIG.  68.  FIG.  69. 


Appearance  of  culture  of  bacillus 
of  tetanus  after  agitating  the  lique- 
fied gelatine.  (Frftnkel  and  Pfeif- 
fer.) 


Six  days'  culture  of  bacillus 
of  tetanus  in  gelatine  (deep 
stab).  (Frftnkel  and  Pfeiffer. 


Agar. — At  brood-heat,  on  agar,  the  growth  is  quite  rapid,  and 
at  the  end  of  forty-eight  hours  gas  bubbles  have  formed  and  the 
growth  nearly  reached  the  surface. 


PATHOGENIC    BACTERIA.  127 

Bouillon. — Adding  glucose  to  the  bouillon  gives  a  medium  in 
which  an  abundant  growth  occurs. 

Staining.— All  the  ordinary  stains,  Grain's  method  also  ;  the 
spores  being  colored  in  the  usual  way. 

Pathogenesis. — A  small  amount  of  the  pure  culture  injected 
under  the  skin  of  experiment  animals  will  cause,  in  two  to  three 
days,  death  from  true  tetanus,  the  tetanic  condition  starting 
from  the  point  of  infection.  At  the  autopsy  nothing  characteristic 
or  abnormal  is  found,  and  the  bacilli  have  disappeared,  except 
near  the  point  of  entrance.  This  fact  is  explained  as  follows  : 

Several  toxic  products  have  been  obtained  from  the  cultures, 
and  they  are  produced  in  the  body,  and  give  rise  to  the  morbid 
symptoms.  These  have  been  isolated,  and  when  injected  singly 
cause  some  of  the  tetanic  symptoms. 

Four  ptomaines  among  them  :  tetanin,  tetanotoxin,  and  spas- 
motoxin  ;  also  a  toxalbumen. 

Immunity. — Kitasato,  by  inoculation  of  sterilized  cultures, 
has  been  able  to  cause  immunity  from  the  effects  of  virulent 
bacilli. 

Habitat. — The  bacillus  is  present  in  garden  earth,  in  manure  ; 
and  even  from  mortar  it  has  been  isolated. 

The  earth  of  special  districts  seems  to  contain  the  bacilli  in 
greater  quantities  than  in  others. 

Bacillus  (Edematis  Maligni.    (Koch.) 

Vibrion  Septique. — ( Pasteur. ) 

Origin. — In  garden  earth,  found  lately  also  in  man,  in  severe 
wounds  when  gangrene  with  osdema  had  developed.  Identical 
with  the  bacillus  found  in  Pasteur's  septicaemia. 

Form. — Bods  somewhat  smaller  than  the  anthrax  bacilli,  the 
ends  rounded  very  sharply.  Long  threads  are  formed.  Very 
large  spores  which  cause  the  rods. to  become  spindle-  or  drum- 
stick-shaped. 

Properties. — Very  motile  ;  liquefy  gelatine  ;  do  not  produce 
any  foul  gaseous  products  in  the  body. 

Growth. — Grows  rapidly,  but  only  when  the  air  is  excluded, 
and  best  at  brood  or  body  heat. 

Roll  Cultures. — (After  Esmarch's  method.)  Small,  round 
glancing  colonies  with  fluid  contents,  under  low  power,  a  mass 


128 


ESSENTIALS    OF    B A CTERIOLOd Y . 


of  motile  threads  in  the  centre,  and  at  the  edges  a  wreath-like 
border. 


FIG.  70. 


FIG.  71. 


Cultures  in  agar  of  malignant 
Oedema,  after  24  hours,  at  37°  C. 
(Frftnkel  and  Pfeiffer.) 


Culture  in  gelatine  of  malig- 
nant (Edema.  (Frftnkel  and 
Pfeiffer.) 


High  Stab  Culture.— With  glucose  gelatine,  the  growth  at  first 
seen  in  the  bottom  of  the  tube,  with  a  general  liquefaction  of 
the  gelatine,  gases  develop  and  a  somewhat  unpleasant  odor. 


PATHOGENIC    BACTERIA.  129 

Agar. — The  gases  develop  more  strongly  in  this  medium,  and 
the  odor  is  more  prominent. 

Guinea-Pig  Bouillon. — In  an  atmosphere  of  hydrogen  cloud- 
ing of  the  entire  culture  medium  without  any  flocculent  pre- 
cipitate until  third  day. 

Staining. — Arc  stained  with  the  ordinary  dyes,  but  Gram's 
method  is  not  applicable. 

Pathogenesis. — When  experiment  animals,  mice  or  guinea- 
pigs,  are  injected  with  a  pure  culture  under  the  skin  they  die  in 
8  to  15  hours,  and  the  following  picture  presents  itself  at  the 
autopsy  :  In  guinea-pigs  from  the  point  of  infection,  spreading 
over  a  large  area,  an  oedema  of  the  subcutaneous  tissues  and 
muscles,  which  are  covered  and  saturated  with  a  clear  red 
serous  exudate  free  from  smell.  This  contains  great  quantities 
of  bacilli. 

The  spleen  is  enlarged,  especially  in  mice.  The  bacilli  are 
not  found  in  the  viscera,  but  are  present  in  great  numbers  on 
the  surface,  i.  6.,  in  the  serous  coverings  of  the  different  organs  ; 
though  when  any  length  of  time  has  elapsed  between  the  death 
of  the  animal  and  the  examination,  they  can  be  found  in  the 
inner  portions  of  the  organs,  for  they  grow  well  upon  the  dead 
bod}'.  In  man  they  have  been  found  in  rapidly  spreading  gan- 
grene. They  are  present  in  the  soil,  in  putrefactions  of  various 
kinds,  and  in  dirty  water. 

Immunity.— IK  produced  by  injection  of  the  sterilized  cul- 
tures, and  also  the  filtered  bloody  serum  of  animals  dead  with 
the  disease. 

Spirillum  of  Relapsing  Fever.    (Obermaier.) 

Syn.  Spirocheete  Obermaieri. 

Origin. —  Found  in  the  blood  of  recurrent  fever  patients, 
described  in  1873. 

Form.— Long,  wavy  threads  (16  to  40  ^  long),  a  true  spiril- 
lum ;  flagella  are  present. 

Properties. — Very  motile.     Has  not  been  cultivated. 

Staining. — Ordinary  aniline  stains.  Bismark  brown  best  for 
tissue  sections. 

Pathogenesis.  —Found  in  the  organs  and  blood  of  recurrent 
fever.  Man  and  monkeys  inoculated  with  blood  from  one  suf- 


loO       ESSENTIALS  OF  BACTERIOLOGY. 

fering  from  this  disease  become  attacked  with  the  fever,  and  in 
their  blood  the  spirillum  is  again  found.  It  is  found  in  the 
blood,  only  in  the  relapses  (during  the  fever).  After  the  attack 
the  spirilla  gather  in  the  spleen  and  gradually  die  there.  It 
has  been  found  in  the  brain,  spleen,  liver,  and  kidneys.  In  the 
secretions  it  has  not  been  discovered. 

Bacillus  Malariae.    (Klebs  and  Tommaci-Crudcli.) 

Origin. — These  two  observers  have  found  a  germ  present  in 
malarial  persons  in  the  blood,  which  produced  an  intermittent 
fever  in  animals  which  had  been  inoculated  with  such  blood. 
They  were  also  found  in  the  soil  of  the  Roman  Campagna. 
Very  little  importance  is  at  present  attached  to  this  germ,  but 
at  the  time  of  its  discovery,  1879,  it  was  thought  to  be  the  cause 
of  malaria. 

Hsematozoa  of  Malaria.  Certain  micro-organisms  are  found 
in  the  blood  of  persons  suffering  from  malaria,  and  have  lately 
been  very  carefully  studied.  They  do  not  belong  to  bacteria, 
being  really  of  animal  origin,  among  the  protozoa;  but  because 
they  are  described  in  the  larger  works  on  bacteria,  it  is  neces- 
sary that  they  be  considered  here. 

Synonyms.  IJamutomonas  Malarice  (Osier).  Plasmodium  Ma- 
larice  (Laveran). 

Form. — Various  shapes  have  been  described,  and  whether 
they  are  all  of  one  micro-organism  or  several  distinct  organisms 
is  not  yet  definitely  settled.  In  the  cell  they  have  been  found 
intra-corpuscular,  and  outside  of  the  cell  extra-corpuscular.  Three 
varieties  of  the  intra-corpuscular  have  been  noted. 

1st.  A  kind  without  pigment,  and  having  amoeboid  movements, 
occupying  about  one-third  of  the  red  corpuscle.  It  is  probably 
the  first  stage  of  the  organism. 

2d.  Pigmented  amvcboid  variety.  The  pigment  probably  ob- 
tained from  the  blood-corpuscle,  which  is  faded  in  color  ;  more 
than  one  may  be  present  in  the  same  corpuscle. 

3d.  The  pigment  set  free  in  grains,  or  surrounded  by  large 
homogeneous  bodies  which  constantly  change  their  outline. 

The  extra-corpuscular  bodies  present  several  peculiar  forms, 
and  are  supposed  to  be  derived  from  the  intra-corpuscular  ones. 

1st.   Tlie  scmilunar  bodies  rf  Laveran ;  spherical  and  crescent- 


PATHOGENIC    BACTERIA. 


131 


shaped  and  motionless,  containing  within  them  little  particles  of 
pigment,  arranged  in  the  centre  and  having  movement.  These 
groups  of  pigment  are  at  times  found  free  in  the  serum. 


FIG.  72. 


.0 


0 


0 


Extra-corpuscular. 


Intra-corpuscular. 
VARIOUS  FORMS  OF  PLASMODIA. 

2d.  Finely  granular  masses  of  protoplasm,  which  assume  vari- 
ous flower-like  shapes,  usually  in  the  form  of  a  rosette  pigment 
in  the  centre  ;  at  the  end  of  a  paroxysm  it  falls  to  pieces,  the 
pigment  being  then  set  free. 

3d.  Oml  bodies  nearly  the  size  of  a  red  corpuscle,  with  long 
motile  flagella  (Carter). 

Cultivation  of  these  organisms  has  not  yet  been  attained. 

Staining  and  Examination  of  Blood.  —Take  the  blood  of  a  per 
son  subject  to  malarial  fevers,  just  before  a  paroxysm.  Having 
first  carefully  cleansed  the  finger,  a  ligature  is  applied,  arid  the 


132       ESSENTIALS  OF  BACTERIOLOGY. 

drop  of  blood  drawn  with  a  needle,  brought  on  a  well-cleaned 
cover-glass,  and  immediately  covered  with  a  second  cover- 
glass.  This  is  now  examined  with  a  strong  objective  (dry  sys- 
tem) by  day-light. 

If,  now,  a  stained  preparation  is  wanted,  the  cover-glasses  are 
slid  apart,  passed  three  times  through  the  flame,  and  a  concen- 
trated solution  of  methylin-blue  left  on  for  a  few  minutes. 

Still  better  is  it  to  allow  a  drop  of  methyliu-blue  solution  in  a 
little  ascitic  fluid  to  flow  slowly  on  the  cover-glass  before  the  blood 
has  become  dry.  The  finer  structure  will  then  be  more  plainly 
brought  out.  Laveran  recommends  the  strong  objective  of  the 
dry  system  for  examining. 

Pathogenesis. — These  organisms  have  been  found  only  in 
malarial  diseases,  and  they  have  been  constantly  found.  Malarial 
paroxysms  have  been  produced  in  a  healthy  person  by  inocu- 
lation of  blood  containing  such  organisms.  They  disappear 
under  the  use  of  quinine. 

Golgi  finds  certain  types  constant  in  tertian,  and  others  again 
peculiar  to  quartan. 

Some,  however,  hold  all  these  various  forms  as  nothing  more 
than  changed  blood-corpuscles.  The  impossibility  of  obtaining 
a  pure  culture  leaves  the  question  still  in  doubt. 

Grassi  and  Fektti  claim  to  have  produced  in  sparrows  and 
man,  by  injection  of  the  blood  of  malarial  persons,  malarial 
fever,  and  found  the  specific  parasites  for  the  different  forms. 
The  amoeboid  or  intra-corpuscular  give  rise  to  the  typical  inter- 
mittent fevers.  In  the  crescent-shaped  extra-corpuscular,  pro- 
ducing the  dumb  ague  or  irregular  fever,  three  different  amoebae 
were  found. 

Hsemoeba  praecox  produces  the  quotidians. 
"          vivax  u         u    tertians, 

malaria?      "         "    quartans. 

They  place  them  with  the  Rhizopoda. 


BACTERIA    PATHOGENIC    FOR    ANIMALS.          133 


CHAPTER  IV. 

BACTERIA  PATHOGENIC  FOR  ANIMALS  BUT  NOT  FOR  MAN. 

Bacillus  of  Symptomatic  Anthrax.    (Bellinger  and  Feser.) 

(Charbon  symptomatique.     Arloing,  Cornevin,  and  Thomas.) 

Origin. — This  bacillus,  described  already  in  1879,  has  only 
lately  been  isolated,  and  by  animal  inoculation  shown  to  be  the 
cause  of  the  "  black-leg"  or  "quarter-evil"  disease  of  cattle. 

Form. — Large  slender  rods,  which  swell  up  at  one  end  or  in 
the  middle  for  the  spore. 

Properties. — They  are  motile,  and  liquefy  gelatine  quite 
rapidly. 

A  rancid  odor  is  developed  in  the  cultures. 

Cultures. — The  growth  occurs  slowly,  and  only  in  an  atmo- 
sphere of  hydrogen,  being  very  easily  destroyed  by  oxygen  and 
carbon  dioxide  ;  grows  best  at  blood  heat ;  under  15°  C.  no 
growth. 

Glucose-gelatine. — In  a  few  days  little  round  colonies  develop, 
which,  under  low  power,  show  hairy  processes  around  a  compact 
centre. 

Stab  Cultures  in  full  test  tubes. — The  growth  first  in  the  lower 
portion  of  the  tube  not  very  characteristic.  Gases  develop 
after  a  few  days,  and  the  gelatine  becomes  liquid. 

Agar  at  brood  temperature,  in  24  to  48  hours,  an  abundant 
growth  with  a  sour  odor  and  abundant  gas  formation. 

Staining.—  Ordinary  methods.  Gram's  method  is  not  appli- 
cable to  the  rods ;  but  the  spores  can  be  colored  by  the  regular 
double  stain  for  spores. 

Pathogenesis. — If  a  small  amount  of  the  culture  be  injected 
under  the  skin  of  a  guinea-pig,  in  twenty  hours  a  rise  of  tempera- 
ture, pain  at  the  site  of  injection,  and  in  a  few  hours  more 
death.  At  the  autopsy,  the  tissues  blackened  in  color  and 
soaked  with  a  bloody  serous  fluid  ;  in  the  connective  tissue  large 
collections  of  gas,  but  only  in  the  neighborhood  of  the  point 


134  ESSENTIALS    OF    BACTERIOLOGY. 

of  infection.  The  bacilli  are  found  in  great  numbers  in  the 
serum,  but  only  appear  in  the  viscera  some  time  after  death, 
when  spores  have  developed. 

The  animals  are  usually  infected  through  wounds  on  the 
extremities  ;  the  stalls  or  meadows  having  been  dirtied  by  the 
spore-containing  blood  of  animals  previously  dead  of  the  dis- 
ease. "  Rauschbrand"  is  the  German  name;  "  Charbon  synip- 
tomatique^  the  French,  from  the  resemblance  in  its  symptoms 
to  anthrax. 

Immunity. — Rabbits,  dogs,  pigs,  and  fowls  are  immune  by 
nature,  but  if  the  bacilli  are  placed  in  a  20  per  cent,  solution  of 
lactic  acid,  and  the  mixture  injected,  the  disease  develops  in 
them.  The  lactic  acid  is  supposed  to  destroy  some  of  the 
natural  resistance  of  the  animal's  cells. 

When  a  bouillon  culture  is  allowed  to  stand  a  few  days,  the 
bacilli  therein  lose  their  virulence,  and  animals  are  no  longer  af- 
fected by  them. 

But  if  they  are  placed  in  20  per  cent,  lactic  add  and  the  mix- 
ture injected,  their  virulence  returns. 

Immunity  is  produced  by  the  injections  of  these  weakened 
cultures,  and  also  by  some  of  the  products  which  have  been  ob- 
tained from  the  cultures. 

Bacillus  of  Chicken  Cholera.    ( Pasteur. ) 

Syn.—Micrococcus  cholera  gallinarum.  Microbe  en  huit.  Ba- 
cillus avitidus.  Bacillus  of  fowl  septicaemia. 

Origin. — In    1879   Perroncito   observed   this 
FIG.  73.  cocci-like  bacillus  in  diseases  of  chickens,  and 

*>     *°  Pasteur,  in  18SO,  isolated  and  reproduced  the 

*<^     '  disease  with  the  microbe  in  question. 

Form. — At  first  it  was  thought  to  be  a  micro- 
coccus,  but  it  has  been  seen  to  be  a  short  rod 
Chicken  cholera      auout  twice  as  long  as  it  is  broad,  the  ends 
bacillus.  slightly  rounded.     The  centre  is  very  slightly 

influenced  by  the  aniline  colors,  the  poles 
easily,  so  that  in  stained  specimens  the  bacillus  looks  like  a 
dumb-bell  or  a  figure-of-eight.  (Microbe  en  huit.) 

Properties. — They  do  not  possess  self-movement ;  do  not 
liquefy  gelatine. 


BACTERIA    PATHOGENIC    FOR    ANIMALS.          135 

Growth. — Occurs  at  ordinary  temperature,  requiring  oxygen 
for  development.  It  grows  very  slowly. 

Gelatine  Plates. — In  the  course  of  three  days  little  round, 
white  colonies,  which  seldom  increase  in  size,  having  a  rough 
border  and  very  finely  granulated. 

Stab  Culture. — A  very  delicate  gray  line  along  the  needle- 
track,  which  does  not  become  much  larger. 

Agar  Stroke  Culture.— A  moist,  grayish-colored  skin,  more 
appreciable  at  brood  heat. 

Potato.— At  brood  heat  after  several  days  a  very  thin,  trans- 
parent growth. 

Staining. — Methyliu  blue  gives  the  best  picture.  Gram's 
method  is  not  applicable.  As  the  bacillus  is  easily  decolorized, 
aniline  oil  is  used  for  dehydrating  tissue  sections,  instead  of 
alcohol. 

Method  : 

Loffler's  methylin  blue  .  £  hour. 

Alcohol 5  seconds. 

Aniline  oil 5  minutes. 

Turpentine 1  minute. 

Xylol  and  Canada  balsam. 

Pathogenesis.— Feeding  the  fowls  or  injecting  under  the  skin 
will  cause  their  death  in  from  12  to  24  hours,  the  symptoms  pre- 
ceding death  being  those  of  a  heavy  septicaemia. 

The  bacillus  is  then  found  in  the  blood  and  viscera,  and  the 
intestinal  discharges,  the  intestines  presenting  a  hemorrhagic 
inflammation. 

Guinea-pigs  and  sheep  do  not  react.  Mice  and  rabbits  are 
affected  in  the  same  manner  as  the  fowls. 

Immunity. — Pasteur,  by  injecting  different-aged  cultures  into 
fowls,  produced  in  them  only  a  local  inflammation,  and  they 
were  then  immune.  But  as  the  strength  of  these  cultures  could 
not  be  estimated,  many  fowls  died  and  the  healthy  ones  were 
endangered  from  the  intestinal  excretions,  which  is  the  chief 
manner  of  infection  naturally  ;  the  fteces  becoming  mixed  with 
the  food. 


136        ESSENTIALS  OF  BACTERIOLOGY. 

Bacteria  of  Hemorrhagic  Septicaemia.    (Hueppe.) 

Under  this  heading  Hueppe  has  gathered  a  number  of  bac- 
teria very  similar  to  the  bacillus  of  chicken  cholera,  differing 
from  it  and  each  other  but  very  little.  They  have  been  described 
by  various  observers  and  found  in  different  diseases. 

1st.  Bacillus  of  American  Swine  Plague.  Syn.  Hog  cholera; 
infectious  pneumonia.  (Billings,  Detmers,  and  Salmon.) 

2d.  Bacillus  of  Swedish-Danish  Swine  Plague.  Swine  </////<- 
theria.  (Selander. ) 

3d.  Bacillus  of  Swine  Plague.    (Loffler,  Scliutz.) 

4th.  Bacillus  of  Texas  Fever.    Cattle  plague.     (Billings.) 

5th.  Bacillus  of  the  Steer  Plague.    (Oreste  and  Armanni.) 

6th.  Bacillus  of  the  French  Swine  Plague.  (Coruil  and 
Chantemesse.) 

7th.  Bacillus  Cuniculicida.    Rabbit  septicaemia.     (Gaffky.) 

8th.  Bacillus  of  Duck  Cholera.    (Cornil  and  Toupet;) 

Differences.  The  various  bacilli  found  in  swine  plague  differ 
from  each  other  very  slightly. 

They  are  but  little  active  in/ourfs,  very  fatal  in  pigs  and  mice. 

In  pigs,  hemorrhages  occur  in  all  the  viscera,  the  mucous  mem- 
brane of  the  intestines  being  ulcerated  and  necrosed. 

The  cultures  of  the  Swedish  swine  plague— No.  2  of  the  above 
list— resemble  the  typhoid  bacillus.  The  form  and  cultures  of 
the  others  are  hardly  to  be  distinguished  from  those  of  the 
chicken  cholera. 

The  bacillus  of  duck  cholera  (No.  8)  is  harmless  for  chickens, 
but  fatal  to  ducks  ;  otherwise  identical  with  chicken  cholera. 

Bacillus  of  Rabbit  Septicaemia.  Similar  in  action  to  the 
chicken  cholera. 

The  other  properties  are  identical  with  those  of  the  microbe 
of  chicken  cholera. 

Bacillus  of  Erysipelas  of  Swine.  (Loffler,  Schiitz. )  Sch  m  ;,„  - 
rollaufbacillus  (German).  Rouget  du pore  (French). 

Origin. — Found  in  the  spleen  of  an  erysipelatous  swine  by 
Loffler  in  1885. 

Form. — One  of  the  smallest  forms  of  bacilli  known  ;  very  thin, 
seldom  longer  than  1  /*,  looking  at  first  like  little  needle-like 
crystals.  Spores  have  not  been  found. 


BACTERIA    PATHOGENIC    FOR    ANIMALS. 


137 


Properties. — They  are  motile  ;  do  not  liquefy  gelatine. 

Growth  .in  culture  at  ordinary  temperature,  very  slowly,  and 
the  less  oxygen  the  better  the  growth. 

Gelatine  Plate. — On  third  day  little  silver-gray  specks,  seen 
best  with  a  dark  background,  coalescing  after  awhile,  pro- 
ducing a  clouding  of  the  entire  plate. 

Stab  Cultures. — In  a  few  days  a -very  light,  silvery -like  clouding, 
which  gradually  involves  the  entire  gelatine  ;  held  up  against 
a  dark  object,  it  comes  plainly  into  view.  , 

Staining. — All  ordinary  dyes  and  Gram's  method  also. 

Tissue  sections  stained  by  Gram's  method  show  the  bacilli  in 
the  cells,  capillaries,  and  arterioles  in  great  numbers. 

Pathogenesis.  —Swine,  mice,  rabbits,  and  pigeons  are  sus- 
ceptible ;  guinea-pigs  and  chickens,  immune. 

When  swine  are  infected  through  food  or  by  injection  a  tor- 
pidity develops  with  diarrhoaa  and  fever,  and  on  the  belly  and 
breast  red  spots  occur  which  coalesce,  but  do  not  give  rise  to 
any  pain  or  swelling.  The  animal  dies  from  exhaustion  in  24  to 
48  hours.  In  mice  the  lids  are  glued  together  with  pus. 

FIG.  74. 


Bacillus  Murisepticus. 

At  the  autopsy  the  liver,  spleen,  and  glands  are  enlarged  and 
congested,  little  hemorrhages  occurring  in  the  intestinal  mucous 
membrane  and  that  of  the  stomach. 


138  ESSENTIALS    OF    13  ACf  ERIOLOG  Y  . 

Bacilli  are  found  in  the  blood  and  all  the  viscera. 

One  attack,  if  withstood,  protects  against  succeeding  ones. 

Immunity. — Has  also  been  attained  by  injecting  vaccines  of 
t\vo  separate  strengths. 

Bacillus  Murisepticus.     (Koch.)    Mouse  septicaemia. 

Origin. — Found  in  the  body  of  a  mouse  which  had  died  from 
injection  of  putrid  blood,  and  described  by  Koch  in  1878. 

Form. — Differs  in  no  particular  from  the  bacillus  of  swine 
erysipelas,  excepting  that  it  is  a  very  little  shorter,  making  it 
the  smallest  known  bacillus.  Spores  have  been  found,  the  cul- 
tures exactly  similar  to  those  of  swine  erysipelas. 

The  pathological  actions  are  also  similar.  Field  mice  are 
immune  ;  whereas  for  house  and  white  mice  the  bacillus  is  fatal 
in  two  to  three  days. 

Micrococcus  of  Mai  de  Pis.  (Nocard.)  Gangrenous  mastitis 
of  sheep. 

Origin. — In  the  milk  and  serum  of  a  sheep  sick  with  the 
"mat  depis." 

Form. — Very  small  cocci  seldom  in  chains. 

Properties,  immotile  ;  liquefying  gelatine. 

Growth.— Growth  occurs  best  between  20°  and  37°  C.,  is  very 
rapid,  and  irrespective  of  ox}'gen. 

Plates  of  Gelatine. — White  round  colonies,  some  on  the  surface 
and  some  in  the  deeper  strata,  with  low  power,  appearing  brown 
surrounded  by  a  transparent  areola. 

Stab  Cwture. — Very  profuse  along  the  needle-track,  in  the 
form  of  a  cone  after  two  days,  the  colonies  having  gathered  at 
the  apex. 

Potato. — A  dirty  gray,  not  very  abundant,  layer  somewhat 
viscid. 

Staining,  with  ordinary  methods  ;  also  Gram's  method. 

PatJwgenesis. — If  a  pure  culture  is  injected  into  the  mammary 
gland  of  sheep,  a  "  mat  de  pis"  is  produced  which  causes  the 
death  of  the  animal  in  24  to  48  hours.  The  breast  is  found 
oedematous,  likewise  the  thighs  and  perineum  ;  the  mammae 
very  much  enlarged,  and  at  the  nipples  a  blue-violet  coloration. 
The  spleen  is  small  and  black  ;  other  animals  are  less  susceptible. 


BACTERIA    PATHOGENIC    FOR    ANIMALS.          139 

In  rabbits  abscesses  at  the  point  of  infection,  but  no  general 
affection. 

Bacillus  Alvei.  (Cheshire  and  Cheyne.)  Bacillus  melittoph- 
tharus.  (Colm.) 

Origin.— In  foul-brood  of  bees. 

.Form.— Slender  rods,  with  round  and  conical-pointed  ends; 
very  large  oval  spores,  the  rod  becoming  spindle-shaped  when 
they  appear. 

Properties. — Motile,  liquefying  gelatine  rapidly. 

Growth. — Grows  best  between  20°  C.  and  37°  C.,  very  slowly  ; 
cerobic. 

Gelatine  Plates. — Small  grooves  are  slowly  formed,  which  unite 
so  as  to  form  a  circle  or  pear-shaped  growth,  from  which  linear 
grooves  again  start. 

Stab  Culture.— Grows  first  on  surface,  then  gradually  along 
the  needle-track,  long  processes  shooting  out  from  the  same, 
clouding  the  gelatine.  Later,  air-bubbles  form  like  the  cholera 
culture,  and  in  two  weeks  the  whole  gelatine  liquefied. 

Staining.  —Do  not  take  aniline  dyes  very  well.  Gram's  method 
is,  however,  applicable. 

Pathogenesis. — If  a  pure  culture  is  spread  over  the  honey- 
comb containing  bee  larvse,  or  if  bees  are  fed  upon  infected 
material,  foul-brood  disease  will  occur.  Mice,  if  injected,  die  in  a 
few  hours.  (Edema  around  the  point  of  infection,  and  many 
bacilli  contained  in  the  cedematous  fluid,  otherwise  no  changes. 

Micrococcus  Amylivorus.    (Bun-ill.) 

Origin.— In  the  disease  called  "Blight,"  which  affects  pear- 
trees  and  other  plants. 

Form. — Small  oval  cells,  never  in  chains,  more  the  form  of  a 
bacillus. 

Pathogenesis. — Introduced  into  small  incisions  in  the  bark  of 
pear-trees  the  trees  perished  from  the  "  blight."  The  starch  of 
the  plant  cell  was  converted  into  carbon  dioxide,  hydrogen, 
and  butyric  acid. 

Bacterium  Termo.    (Cohn.) 

This  was  a  name  given  to  a  form  of  micro-organism  found  in 
decomposing  albuminous  material,  and  was  supposed  to  be  one 
specific  germ.  Hauser,  in  1885,  found  three  different  distinct 


140       ESSENTIALS  OF  BACTERIOLOGY. 

microbes  which  he  grouped  under  the  common  name  of  Protons, 
which  have  the  putrefying  properties  ascribed  to  B.  Termo. 

Proteus  Vulgaris. 

Origin. — In  putrid  animal  matter,  in  meconium  and  in  water. 

Form. — Small  rods,  slightly  curved,  of  varying  lengths,  often 
in  twisted  chains,  having  long  cilia  or  flagella. 

Properties. — Very  motile,  and  very  soon  liquefying  gelatine  ; 
forms  hydrogen  sulphide  gas  ;  causes  putrefaction  in  meat. 

Growth. — Growth  very  rapid,  best  at  24°  C.,  is  facultative 
aerobic. 

Gelatine  Plates. — Yellowish-brown,  irregular  colonies,  with 
prolongations  in  every  direction,  forming  all  sorts  of  figures  ;  an 
impression  preparation  shows  these  spider-leg  processes  to  con- 
sist of  bacilli  in  regular  order. 

Stab  Culture. — The  gelatine  soon  liquid,  a  gray  layer  on  the 
surface,  but  the  chief  part  of  the  culture  in  small  crumbs  at  the 
bottom. 

Pathogenesis. — Rabbits  and  guinea-pigs  injected  subcutane- 
ously  die  quickly,  a  form  of  toxaemia,  hemorrhagic  condition  of 
lungs  and  intestines  present.  When  neurin  is  injected  previ- 
ously the  animals  do  not  die.  This  ptomaine  is  supposed  to  be 
generated  by  the  proteus  vulgaris. 

Proteus  Mirabilis.     (Hauser.) 

Differs  from  P.  vulgaris  in  that  the  gelatine  is  less  rapidly 
liquefied.  Found  also  in  putrid  material. 

Proteus  Zenkeri.     (Hauser.) 

Does  not  liquefy  gelatine  ;  otherwise  similar  to  the  other  two. 

We  have  now  considered  some  of  the  characteristics  of  the 
more  important  bacteria.  The  scope  of  this  work  does  not  allow 
a  more  extended  study  than  we  have  made,  which,  as  we  are 
aware,  has  been  very  superficial.  The  larger  works  must  be 
referred  to,  if  a  deeper  interest  is  taken  in  the  subject. 


APPENDIX. 


YEASTS  AND  MOULDS. 

IN  works  on  bacteria,  these  true  fungi,  yeasts  and  moulds,  are 
usually  considered.  They  are  so  closely  related  to  bacteria,  and 
so  often  contaminate  the  culture  media,  and  are  so  similar  in 
many  respects,  that  a  description  is  almost  a  necessity. 

But  there  are  several  thousand  varieties,  and  we  cannot 
attempt  to  describe  even  all  of  the  more  important  ones.  It 
will  answer  our  purpose  to  detail  a  few  of  the  more  common 
kinds,  and  give  the  principal  features  of  the  different  orders. 

Fungi  exist  without  chloropliyl. 

Sacchuromycetes  or  Yeasts  increase  through  budding ;  the 
spores  attached  to  the  mother  cell  like  a  tuber  on  a  potato. 

Yeasts  are  the  cause  of  alcoholic  fermentation  in  the  saccha- 
roses. A  description  of  the  most  common  ones  will  suffice. 

Saccharomyces  Cerevisiae.  (Torula  Cerevisice.)  This  is  the 
ordinary  beer  yeast. 

Form. — Kound  and  oval  cells  ;  a  thin  membrane  inclosing  a 
granular  mass,  in  which  usually  can  be  seen  three  or  four  irre- 
gular-shaped spores.  When  these  become  full  grown  they  pass 
through  the  cell  wall  and  form  a  daughter  cell.  Sometimes  long 
chains  are  produced  by  the  attached  daughter  cells. 

Growth.—  They  can  be  cultivated  as  bacteria  in  bouillon,  but 
they  grow  best  in  beer. 

There  are  several  varieties  of  beer  yeast,  each  one  giving  a 
characteristic  taste  to  the  beer.  Brewers,  by  paying  special 
attention  to  the  nutrient  media,  cultivate  yeasts  which  give  to 
their  beers  individual  flavors. 

Mixed  yeast  gives  rise  to  a  poor  quality  of  beer. 

Saccharomyces  Rosaceus.  S.  Niger  and  S.  Albicans.  These 
yeasts  are  found  in  the  air  ;  and  instead  of  producing  alcoholic 

(141) 


142  APPENDIX. 

fermentation  they  give  rise  to  a  pigment  in  the  culture  media. 
They  grow  upon  gelatine  which  they  do  not  liquefy. 

Saccharomyces  Mycoderma.  This  yeast  forms  a  mould-like 
growth,  a  skin,  on  the  surface  of  fermented  liquids,  but  does  not 
cause  any  fermentation  itself.  It  forms  the  common  "mould" 
on  wine,  preserves,  and  "sour  krout." 

Oidium.  A  form  which  seems  to  be  the  bridge  between  the 
yeast  and  the  moulds  is  the  oidium.  Sometimes  it  resembles 
the  yeasts,  sometimes  the  moulds,  and  often  both  forms  are 
found  in  the  same  culture.  Several  are  pathogenic  for  man. 

Oidium  Lactis. 

Origin. — In  sour  milk  and  butter. 

Form. — The  branches  or  hyphens  break  up  into  short  rod-like 
spores.  No  sporangium,  as  in  moulds. 

Growth.  — In  milk  it  appears  as  a  white  mould. 

Artificially  cultured  on  gelatine  plates,  or  milk  gelatine  plates, 
it  forms  satin-like,  star-shaped  colonies,  which  slowly  liquefy. 
Under  microscope  the  form  of  the  fungus  is  well  seen. 

Ayar  Stroke  Culture. — The  little  stars,  very  nicely  seen  at  first ; 
then  the  culture  becomes  covered  with  them,  causing  a  smeared 
layer  to  appear  over  the  whole  surface,  with  a  sour  odor. 

Properties. — The  milk  is  not  changed  in  any  special  way.  It 
is  not  pathogenic  for  man  or  animals.  It  is  found  when  the 
milk  begins  to  sour. 

Oidium  Albicans.    (Soor.}     Thrush  Fungus. 

Origin.— Mucous  membrane  of  the  mouth,  especially  of  infants. 

Form. — Taken  from  the  surface  of  the  culture,  a  form  like 
yeasts  ;  but  in  the  deeper  layers,  mycelia  with  hyphens  occurs. 

Growth.— Not  liquefying ;  snow-white  colonies  on  gelatine 
plates. 

Stab  CuHure. — Radiating  yellow  or  white  processes  spring  from 
the  line  made  by  the  needle,  those  near  the  surface  having  oval 
ends. 

Potatoes. — The  yeast  form,  develops  as  thick  white  colonies. 

Bread  Mash. — Snow-white  veil  over  the  surface. 

Pathogenesis.—ln  man  the  parasitic  thrush,  or  "  white  mouth," 
is  caused  by  this  fungus.  In  the  white  patches  the  spores  and 
filaments  of  this  microbe  can  be  found.  Rabbits  receiving  an 


YEASTS    AND    MOULDS.  143 

intravenous  injection  perish  in  twenty-four  to  forty-eight  hours, 
the  viscera  being  filled  with  mycelia. 

True  Moulds.  Fliigge  has  made  five  distinct  divisions  of 
moulds.  It  will,  however,  serve  our  purpose  to  classify  those 
to  be  described  under  three  headings :  Pendllium,  Mucor,  and 
Aspergillus. 

Pencillium  Glaucum. 

Origin. — The  most  widely  distributed  of  all  moulds,  found 
wherever  moulds  can  exist. 

Form. — From  the  mycelium,  hyphens  spring  which  divide  into 
basidia  (branches),  from  which  tiny  filaments  arise  (sterygmata), 
arranged  like  a  brush  or  tuft.  On  each  sterygma  a  little  bead 
or  conidium  forms,  which  is  the  spore.  In  this  particular  fungus 
the  spores  in  mass  appear  green. 

Growth.— It  develops  only  at  ordinary  temperatures,  forming 
thick  grayish-green  moulds  on  bread-mash.  At  first  these  ap- 
pear white,  but  as  soon  as  the  spores  form,  the  green  predomi- 
nates. Gelatine  is  liquefied  by  it. 

Mucor  Mucedo.  Next  to  the  pencillium  glaucum,  this  is  the 
most  common  mould.  Found  in  horse  dung,  in  nuts,  and 
apples,  in  bread  and  potatoes  as  a  white  mould. 

Form.— The  mycelium  sends  out  several  branches,  on  one  of 
which  a  pointed  stem  is  formed  which  enlarges  to  form  a  globu- 
lar head,  a  spore-bulb,  or  Sporangium..  The  spore-bulb  is  par- 
titioned onMnto  cells  in  which  large  oval  spores  lie.  When  the 
spores  are  ripe  a  cap  forms  around  the  bulb,  the  walls  break 
down  and  the  wind  scatters  the  spores,  leaving  the  cap  or 
"  ccilumella"  behind. 

Growth. — Takes  place  at  higher  temperatures  on  acid  media. 

It  is  not  Pathogenic. 

Achorion  Schonleinii. 

Tricophyton  Tonsurans. 

Microsporon  Furfur. 

These  three  forms  are  similar  to  each  other  in  nearly  every 
particular  and  resemble  in  some  respects  the  oidium  lactis,  in 
other  ways  the  mucors.  The  first  one,  Achorion  Schb'nleinii,  was 
discovered  by  Schonlein  in  1839,  in  Favus,  and  is  now  known  as 
the  direct  cause  of  this  skin  disease. 


144  APPENDIX. 

Origin. — Found  in  the  scaly  crusts. 

Form. — Similar  to  oidiuiu  lactis.. 

Growth. — Is  very  sparse.  On  gelatine  round  white  masses 
inclosed  by  a  zone  of  liquefied  gelatine. 

In  milk  it  is  destroyed. 

Pathoyenesis. — Causes  favus  in  man. 

Tricophyton  Tonsurans.    Found,  in  1854,  by  Bazin,  in  Tinea. 

Form. — Similar  to  the  achorion  or  i'avus  fungus. 

Growth.— Somewhat  more  rapid  than  the  favus,  and  the  gela- 
tine quickly  liquefied.  Old  cultures  are  of  an  orange-yellow 
color.  Colonies  have  a  star-shaped  form. 

Patlwyenesis.  —  Herpes  tonsurans  and  the  various  tinea3  are 
produced  by  this  fungus. 

Microsporon  Furfur.  Found  in  tinea  versicolor,  almost  iden- 
tical with  the  above,  forms  dry  yellow  spots,  usually  on  the  chest 
in  persons  suffering  from  wasting  diseases. 

Aspergillus  Glaucus, 

Origin. — In  saccharine  fruits. 

Form. — The  hyphen  has  formed  upon  its  further  end  a  bulb, 
from  which  pear-shaped  sterygmata  arise  and  bear  upon  their 
ends  the  conidia  or  spores. 

Growth.  —  Best  upon  fruit  juices.  Non-pathoyeni\  The  mould 
is  green.  AsperyUhtifiavua  has  the  tufts  and  spores  of  a  yellow 
color. 

A.  Fumigatus.  Is  pathogenic  for  rabbits  when  injected  into 
them.  At  the  autopsy  their  viscera  are  found  filled  with  the 
mould. 

Examination  of  Yeasts  and  Moulds.  Yeasts  and  moulds  are 
best  examined  in  the  unstained  condition.  A  small  portion  of 
the  colony  rubbed  up  with  a  mixture  of  alcohol  and  a  few  drops 
of  liquor  ammonia  ;  of  this,  a  little  is  brought  upon  the  glass- 
slide  covered  with  a  drop  of  glycerine  and  the  cover-glass  pressed 
upon  it.  If  the  preparation  is  to  be  saved,  the  cover-glass  is 
secured  by  sealing-wax  around  the  edges.  Yeasts  take  methy- 
line-blue  stain  very  well. 

Ray  Fungus.  A  division  containing  the  actinomyces.  (Bol- 
lenger  and  Israel.) 

Oriyin.— In  actinomycosis  of  man  and  cattle,  in  the  growth. 


RAY    FUNGUS.  145 

Form. — In  the  pus  or  scrapings,  little  yellow  grains  about 
the  size  of  a  pin's  head  are  seen  by  the  naked  eye.  When  one  of 
these  points  is  flattened  out  between  the  cover-glass  and  slide 
and  placed  under  microscope  (200  x),  aster-shaped  figures  will 

Kir,.   75. 


Actinomyces. 

be  seen,  the  centre  thick,  radiating  from  it,  little  hyphens,  which 
become  thicker  and  rounder  at  their  peripheral  end.  These 
bottle-shaped  hyphens  are  supposed  to  be  the  spore-bearing 
organs.  Some  of  these  may  have  separated  from  the  main  body 
and  lie  loose  or  attached  to  it  by  a  very  thin  filament. 

Growth.— Develops  only  at  brood  heat  and  by  exclusion  of 
oxygen. 

In  Agar. — After  several  weeks  a  yellowish  growth  was  ob- 
tained, but  this  consisted  mainly  of  mycelia,  the  club-shaped  or 
conical  rays  not  forming. 

In  eggs  a  growth  developed  when  the  method  of  Hueppe  was 
carried  out. 

Pathogenesis. — When  a  portion  of  the  growth  obtained  in 
eggs  was  injected  into  the  abdominal  cavity  of  a  rabbit,  actinomy- 
cotic  processes  developed  upon  the  peritoneum. 

It  usually  gains  access  to  the  living  body  through  a  wound  in 
the  gum  or  some  caries  of  the  teeth.  A  new  growth  is  formed, 
ulceration  being  first  set  up. 

The  new  tissue,  composed  of  round  cells,  then  undergoes  soft- 
10 


146  APPENDIX. 

en  ing,  purulent  collections  form  and  the  normal  structure  is 
destroyed. 

The  usual  seat  is  in  the  maxillary  bones,  but  the  fungus  has 
been  found  in  the  lungs,  tonsils,  intestines,  and  various  other 
organs  in  man  and  cattle. 

Examination. — Well  seen  in  the  unstained  condition.  From 
the  pus  or  scraping  a  small  portion  is  taken  and  squeezed  upon 
the  glass  slide  ;  if  calcareous  matter  is  present,  a  drop  of  nitric 
acid  will  dissolve  the  same. 

Glycerine  will  preserve  the  preparation. 

Staining. — Cover-glass  specimens  stained  best  with  Gram's 
method.  Tissue  sections  should  be  stained  as  follows  : — 

Ziebel's  carbol-fuchsin,  ten  minutes.     Rinse  in  water. 

Cone,  alcohol  sol.  of  picric  acid,  li ve  minutes.     Rinse  in  water. 

Alcohol,  50  per  cent. ,  fifteen  minutes.  Alcohol  absolute,  clove 
oil,  balsam. 

The  rays  stained  red,  the  tissue  yellow. 

Examination  of  Air,  Soil,  and  Water. 

Air.— Many  germs  are  constantly  found  in  the  atmosphere 
about  us.  Bacteria  unaided  do  not  rise  into  the  air  and  fly 
about ;  they  usually  become  mixed  with  small  particles  of  dirt 
or  dust  and  are  moved  with  the  wind.  The  more  dust  the  more 
bacteria,  and  therefore  the  air  in  summer  contains  a  greater 
number  than  the  air  in  winter,  and  all  the  other  differences  can 
be  attributed  to  the  greater  or  less  quantity  of  dust  and  wind. 

Methods  of  Examination.  The  simplest  method  is  to  ex- 
pose a  glass  or  dish  covered  with  gelatine  in  a  dust-laden 
atmosphere  or  in  the  place  to  be  examined.  In  the  course  of 
24  to  48  hours  colonies  will  be  seen  formed  wherever  a  germ  has 
fallen.  But  this  method  will  not  give  any  accurate  results  in 
regard  to  the  number  of  bacteria  in  a  given  space  ;  for  such  a 
purpose  somewhat  more  complicated  methods  are  needed,  so 
that  a  certain  amount  of  air  can  come  in  contact  with  the 
culture  media  at  a  certain  regulated  rate  of  speed. 

Hesse's  Method,  This  is  the  most  useful  of  the  various 
methods  in  vogue. 

A  glass  cylinder,  70  centimetres  long  and  3.5  centimetres  in 
diameter,  is  covered  at  one  end,  by  two  rubber  caps,  the  inner 


AIR,    SOIL,    AND    WATER. 


147 


one  having  a  hole  in  its  centre  10  millimetres  in  diameter  ;  and 
at  the  end  B  a  rubber  cork  fits  in  the  cylinder ;  through  this 
cork  a  glass  tube  10  mm.  in  diameter  passes,  which  is  plugged 
at  both  ends  with  cotton.  The  cylinder  and  fittings  are  first 
washed  in  alcohol  and  sublimate  and  then  placed  for  one  hour 
in  the  steam  chamber. 

Removing  the  cork  of  the  cylinder,  50  cubic  centimetres  of 
sterile  gelatine  in  a  fluid  condition  are  introduced  and  rolled 
out  on  the  sides  of  the  tube,  after  the  manner  of  Esmarch, 
leaving  a  somewhat  thicker  coating  along  the  under  side  of  the 

FIG.  76. 


cylinder.  The  eeroscqpe,  as  the  cylinder  and  its  fittings  are 
called,  is  placed  upon  an  ordinary  photographer's  tripod  and 
the  glass  tube,  which  passes  through  the  rubber  cork,  connected 
with  an  aspirator,  the  cotton  having  first  been  removed  from  its 


148 


APPENDIX. 


FIG.  77. 


outer  end.  The  aspirator  consists  of  two  ordinary  wash-bottlrs 
connected  with  each  other  by  a  rubber  tube,  C.  They  are  at- 
tached to  the  tripod  with  a  small  hook  one  above  the  other,  the 
upper  one  half  filled  with  water  and  slightly  tilted. 

When  the  apparatus  is  wanted,  the  outer  rubber 
cap  at  the  end  A  of  the  aeroscope  is  removed,  the 
air  can  then  pass  through  the  small  hole  in  the 
other  cap.  and  the  germs  fall  upon  the  gelatine  in 
the  tube,  the  cotton  in  the  small  glass  tube  at  the 
other  end  preventing  the  germs  from  getting  out. 
The  aspirator  is  set  in  use  by  tilting  the  upper 
bottle  so  that  the  water  flows  into  the  lower,  this 
creates  suction  and  draws  the  air  through  the 
aeroscope. 

The  amount  entering  estimated  by  the  capacity 
of  the  wash-bottle.  The  rate  at  which  it  enters 
depending  upon  the  rate  of  the  flow  of  water, 
which  can  be  regulated. 

Hesse  advises  for  rooms  and  closed  spaces  1  to  5 
litres,  at  the  rate  of  2  minutes  a  litre,  and  for  open 
spaces,  10  to  20  litres  at  4  minutes  a  litre.  Plate 
cultures  can  be  made  from  the  colonies  which  de- 
velop in  8  to  10  days  in  the  cylinder. 

Petri's  Method.  The  air  pumped  or  sucked 
through  sand  filters,  and  the  sand  then  mixed  with 
gelatine. 

Sand  is  sterilized   by  -heating  to  redness,  and 
while  still  warm  placed  in  test  tubes  which  are 
Sand  filter        then   plugged.      (Sand   which    has    been    passed 
through  a  sieve  with  meshes  0.25  millimetres  wide 
is  the  kind  required.)     A  glass  tube  9  centimetres  long  is  pro- 
vided with  two  portions  of  sand  each  3  cm.  long  and  £  cm.  apart, 
little  plates  of  brass  gauze  keeping  the  portions  in  position. 

The  tube  and  its  contents  now  sterilized  in  hot  air  oven  at 
150°  C.,  the  ends  having  first  been  plugged  with  cotton. 

One  end  of  the  tube  is  then  fitted  with  a  rubber  cork  through 
which  passes  a  glass  tube,  which  is  connected  with  an  aspirator 
(a  hand-pump  with  a  known  capacity). 


AIR,    SOIL,    AND    WATER.  149 

If  a  hundred  litres  of  air  pass  through  the  tube  in  fifteen  min- 
utes the  germs  should  all  be  arrested  in  the  first  sand  filter. 

And  when  the  filters  are  removed  and  thoroughly  mixed  with 
gelatine,  each  filter  for  itself,  there  should  be  no  colonies  de- 
veloped from  the  second  filter,  t.  e.,  the  one  nearest  the  aspirator. 

Varieties  Found  in  Air.  The  only  pathogenic  ones  found  with 
any  constancy  are  the  staphylo-coccus  aureus  and  citreus  ;  but 
any  bacterium  can  be,  through  accident,  lifted  into  the  atmo- 
sphere, and  in  certain  places-may  be  always  found— the  bacillus 
tuberculosis,  for  example,  in  rooms  where  many  consumptives 
are  living. 

Non-Pathogenic.  The  micrococci  predominate.  Sarcina, 
yeasts,  and  moulds  constantly  contaminate  cultures. 

In  the  ordinary  habitations  the  average  number  of  germs  to 
the  litre  of  air  does  not  exceed  five. 

Around  water-closets,  where  one  would  imagine  a  great  num- 
ber to  exist,  owing  to  the  undisturbed  condition  of  the  air,  but 
few  will  be  found. 

Examination  of  Water.  The  bacteriological  examination  of 
water  is  to-day  of  as  much  importance  as  the  chemical  analy- 
sis, and  must  go  hand  in  hand  with  it. 

At  the  start  we  must  say  that  a  water  containing  thousands 
of  germs  to  the  cubic  centimeter  is  far  less  dangerous  than  one 
containing  but  2  germs,  if  one  of  these  two  be  a  typhoid  bacil- 
lus. It  is  not  the  number  that  proves  dangerous ;  it  is  the 
kind. 

If  a  natural  water  contains  more  than  500  germs  to  the  cubic 
centimeter,  it  were  well  to  examine  its  source. 

Bacteriology  performs  the  greatest  service  in  testing  the  devices 
which  are  intended  to  render  water  fit  for  drinking. 

As  a  diagnostic  aid  the  examination  is  of  but  little  use.  An 
epidemic  of  typhoid  fever  occurs,  the  water  is  suspected,  an  ex- 
amination is  undertaken  ;  but  the  days  of  incubation  and  the 
days  passed  before  the  water  is  analyzed  have  given  the  typhoid 
germs,  if  any  had  been  present,  ample  time  to  disappear,  since 
in  water  that  contains  other  bacteria  they  live  a  very  short  time 
only.  Again,  the  water  tested  one  day  may  be  entirely  free  and 
the  next  day  contain  a  great  number,  and  before  the  typhoid 


150  APPENDIX. 

germ  can  be  proven  to  be  present  in  tbat  particular  water,  tbe 
epidemic  may  be  pa>t. 

Purity  of  Waters.  Tbe  purest  water  we  have  is  tbe  natural 
spring  water— water  that  has  slowly  filtered  its  way  through 
various  layers  of  gravel  and  sand  and  cornes  finally  clear  and 
sparkling  from  tbe  ground.  It  is  without  germs  ;  but  let  such 
a  water  stand  walled  up  in  cisterns  or  wells,  it  becomes  as 
surface  water,  open  to  all  sorts  of  impurities,  and  the  bacterial 
nature  of  it  changes  every  moment. 

Artesian  or  Driven  Well.  The  driven  well  will  secure  to  a  cer- 
tain extent  a  pure  water.  It  is  the  only  form  of  well  or  cistern 
tbat  will  insure  this,  since  tbe  water  does  not  become  stagnant 
in  it ;  but  it  may  connect  with  an  outhouse,  the  soil  being  very 
loose,  allowing  the  products  of  germs  of  refuse  water  to  find  their 
way  into  the  well.  If  a  chemical  examination  shows  increased 
amounts  of  chloride  of  sodium,  a  contamination  can  be  mooted. 

Filtered  Water.  Dangerous  as  surface  water  is,  the  greater 
quantity  used,  is  such  :  the  inhabitants  of  larger  towns  and  cities 
using  chiefly  tbe  rivers  and  other  large  waters  which  course 
near  them  for  drinking  purposes.  A  purification  or  filtration 
can  in  a  certain  measure  render  these  waters  harmless. 

Filtration  is  often  carried  on  on  a  large  scale  in  the  water- 
works of  cities  and  towns. 

Bacteriological  examination  is  here  of  great  service  to  deter- 
mine if  a  water,  which  has  been  filtered  and  may  have  a  very 
clear  appearance,  and  give  no  harmful  chemical  reaction,  yet 
be  entirely  free,  or  nearly  so,  from  germs ;  in  other  words,  if 
the  filter  is  a  germ  filter  or  not. 

Charcoal  Sponge  and  Asbestos.  The  materinls  formerly  in  use 
are  objectionable  because  germs  readily  develop  on  them  and 
clog  them,  so  that  they  require  frequent  renewal.  In  very 
large  filters,  sand  and  gravel  give  the  best  results ;  tbe  number 
of  germs  in  a  cubic  centimetre  was  reduced  to  forty  or  fifty  and 
kept  at  that  number.  This  is  a  very  pure  water  for  a  city  water, 
though,  as  we  stated  before,  not  a  safe  one,  for  among  those 
forty  germs  very  dangerous  ones  may  be.  It  is  then  necessary 
for  the  users  to  refilter  the  water  before  drinking  it,  through  a 
material  which  will  not  allow  any  germs  to  pass. 


AIR,    SOIL,   AND    WATER.  151 

Pasteur-Chamberland  Filter.  This  very  perfect  filter,  which 
is  now  in  almost  universal  use,  consists  of  a  piece  of  polished 
porcelain  in  the  form  of  a  cylinder  closed  at  one  end  and  pointed 
at  the  other.  It  is  placed  in  another  cylinder  of  glass  or  rubber 
and  the  pointed  portion  connected  with  a  bottle  containing  the 
water,  or  directly  with  faucet  of  the  water-pipe.  The  water 
courses  through  the  porcelain  very  slowly  and  comes  out  entirely 
free  from  germs  ;  pipe-clay,  bisque,  and  kaolin  are  also  perfect 
filters.  The  only  disadvantage  is  the  long  time  it  takes  for  the 
water  to  pass  through.  Pressure  is  used  to  accelerate  the  pas- 
sage in  the  form  of  an  aspirator  or  air-pump.  (See  Fig.  37.) 

The  force  of  the  hydrant  water  is  also  sufficient  to  produce  a 
steady,  small  stream. 

These  porcelain  cylinders  can  easily  be  sterilized  and  the 
pores  washed  out. 

Boiling  as  a  means  of  purifying.  When  such  a  filter  cannot 
be  obtained,  the  only  alternative  is  to  boil  all  the  water  to  be 
used  for  drinking  ;  and  this  should  especially  be  done  in  times 
of  typhoid  and  cholera  epidemics. 

Methods  of  Examination.  Since  the  germs  rapidly  multiply 
in  stagnant  water,  an  examination  must  not  be  delayed  longer 
than  an  hour  after  the  water  has  been  collected.  Every  pre- 
caution must  be  taken  in  the  way  of  cleanliness  to  prevent  con- 
tamination ;  sterilized  flasks,  pipettes,  and  plugs  should,  or  rather 
must,  be  at  hand,  and  the  gelatine  tubes  best  inoculated  on  the 
spot.  If  this  cannot  be  done,  the  sample  should  be  packed  in 
ice  until  it  arrives  at  the  laboratory,  which,  as  before  stated, 
should  not  be  later  than  an  hour  after  collection.  The  sample 
is  placed  in  a  sterilized  glass  flask,  and  the  flask  then  closed  with 
a  sterile  cotton  plug.  A  sterilized  pipette  is  then  dipped  into 
the  flask  and  1  c.c.  of  the  water  withdrawn  in  it  and  added  to 
a  tube  of  gelatine,  the  gelatine  being  in  a  fluid  condition.  To  a 
second  tube,  £  c.c.  is  added.  The  tubes  are  then  shaken  so  as 
to  thoroughly  mix  the  water  with  the  gelatine,  and  then  poured 
upon  wide  glass  plates— one  plate  for  each  tube  ;  the  plates  are 
then  placed  in  the  moist  chamber,  and  in  two  to  three  days 
examined.  If  the  germs  are  equally  divided,  there  should  be 


152  APPENDIX. 

one-half  the  number  on  one  plate  that  there  is  on  the  other  ; 
thus  the  £  c.c.  serves  as  control. 

Water  that  is  very  rich  in  germs  requires  dilution  with  ster- 
ilized water  50  to  100  times. 

To  count  the  colonies  which  develop  upon  the  plates,  a  spe- 
cial apparatus  has  been  designed,  for,  unaided,  the  eye  cannot 
see  them  all. 

Wolfhugel's  Apparatus.  A  glass  plate  divided  into  squares, 
each  a  centimeter  large,  and  some  of  these  subdivided.  This 
plate  is  placed  above  the  gelatine  plate  with  the  colonies,  and 
the  number  in  several  quadrants  taken,  a  lens  being  used  to  see 
the  smaller  ones. 

Varieties  Found.  The  usual  kinds  found  are  non-pathogenic, 
but,  as  is  well  known,  typhoid  and  cholera  are  principally  spread 
through  drinking  water,  and  many  other  germs  may  and  do 
find  their  way  into  the  water.  Many  of  the  common  varieties 
give  rise  to  fluorescence,  or  produce  pigment. 

Eisenberg  gives  100  different  varieties  as  ordinarily  found. 
As  mentioned  before,  2  bacteria  to  a  cubic  centimeter,  one  of 
them  typhoid,  give  more  danger  to  a  water  than  thousands  of 
non-pathogenic  one?.  When,  however,  more  than  200  bacteria 
to  the  c.c.  are  found,  such  a  water  ought  not  to  he  considered 
potable.  Distilled  water  forms  often  a  good  medium  for  some 
bacteria. 

The  Examination  of  the  Soil.  The  upper  layers  of  the  soil 
contain  a  great  many  bacteria,  but  because  of  the  difficulty  in 
analyzing  the  same,  the  results  are  neither  accurate  nor  con- 
stant. The  principal  trouble  lies  in  the  mixing  of  the  earth 
with  the  nutrient  medium  ;  little  particles  of  ground  will  cling 
to  the  walls  of  the  tube,  or  be  imbedded  in  the  gelatine,  and 
may  contain  within  them  myriads  of  bacteria.  As  with  water, 
the  soil  must  be  examined  immediately  or  very  soon  after  it  is 
collected,  the  bacteria  rapidly  multiplying  in  it. 

When  the  deeper  layers  are  to  be  examined,  some  precautions 
must  be  taken  to  avoid  contamination  with  the  other  portions  of 
the  soil.  One  method,  very  laborious  and  not  often  practical,  is 
to  dig  a  hole  near  the  spot  to  be  examined  and  take  the  earth 
from  the  sides  of  this  excavation. 


AIR,    SOIL,    AND    WATER.  153 

Frankel's  Borer.  Eriinkel  has  devised  a  small  apparatus  in 
the  form  of  a  borer,  which  contains  near  its  lower  end  a  small 
cavity,  which  can  be  closed  up  by  turning  the  handle,  or  opened 
by  turning  in  the  opposite  direction. 

It  is  introduced  with  the  cavity  closed,  and  when  it  is  at  the 
desired  depth,  the  handle  is  turned,  the  earth  enters  the  cavity, 
the  handle  again  turned,  incloses  it  completely,  and  the  borer  is 
then  withdrawn. 

The  earth  can  then  be  mixed  with  the  gelatine  in  a  tube,  and 
this  gelatine  then  rolled  on  the  walls  of  the  tube  after  the  man- 
ner of  Esmarch,  or  it  can  be  poured  upon  a  glass  plate,  and  the 
colonies  developed  so. 

Another  method  is  to  wash  the  earth  with  sterilized  water, 
and  the  water  then  mixed  with  the  gelatine,  as  many  of  the 
germs  are  taken  up  by  the  water. 

The  roll-cultures  of  Esmarch  give  the  best  results,  many  of 
the  varieties  usually  found  being  anaerobic. 

Animals  inoculated  with  the  soil  around  Berlin  die  almost 
always  of  malignant  oedema,  and  with  that  of  some  other  towns 
invariably  of  tetanus. 

CONCLUSION.  In  tracing  thus  briefly  the  characteristics  of 
the  more  important  bacteria,  and  the  various  methods  used  in 
studying  them,  we  are  conscious  of  the  very  superficial  manner 
in  which  this  has  been  done.  We  excuse  ourselves,  however, 
on  the  ground  that  this  work  is  but  a  wedge  with  which  to  enter 
upon  the  study,  or,  for  those  who  do  not  care  to  proceed  further, 
an  eminence  from  which  a  fair  view  of  the  ground  can  be  ob- 
tained. In  this,  its  humble  mission,  we  trust  it  may  meet  with 
success. 


INDEX. 


ABBE'S  condenser,  26 
Achorion  Schouleinii,  143 
Actinomyces,  144 
Actinouiycosis,  145 
M robins,  24 

facultative,  24 

obligative,  24 
Agar-agar,  49 

bouillon,  49 

glycerine,  50 
Air,  examination  of,  146 
Anaerobins,  24 

facultative,  24 

obligative,  24 
Aniline  dyes,  30 

oil,  38 

oil  water,  31 

Animals  for  experiment,  68 
Anthrax,  84 
Arthrospores,  18 
Asbestos  filter,  150 
Aspergillus  fumigatus,  144 

glaucus,  144 
Asporogenic  bacteria,  22 
Attenuation,  64 
Autoclave  of  Chamberland,  41 


BACILLUS  acidi  lactici,  76 
alvei,  139 
amylobacter,  77 
anthracis,  84 
avicipis,  134 
butyricus,  77 
capsule,  116 
coeruleus,  79 
comma,  105 
cuniculicida,  136 
erythrosporus,  80 
faeces,  103 
fluorescens,  120 

liquefaciens,  80 
hay,  75 


Bacillus,  indicus,  73 
Klebs-Loffler,  100 
lactis  cyanogenus,  78 

erythogenes,  78 
lepra,  96 
malariae,  130 
mallei,  98 
megaterium,  74 
melittoptharus,  139 
mesentericus  vulgatus,  73 
Milzbrand,  84 
murisepticus,  138 
mycoides,  74 
Neapolitanus,  103,  104 
cedernatus  maligni,  127 
of  American  swine  plague,  136 
of  anthrax,  84 
of  bluish-green  pus,  120 
of  chicken  cholera,  134 
of  diphtheria,  100 
of  duck  cholera.  136 
of  fowl  septicaemia,  134 
of  French  swine  plague,  136 
of  glanders,  98 
of  mouse  septicaemia,  138 
of  rhinoscleroma,  115 
of  steer  plague,  136 
of  Swedish-Danish  plague,  136 
of  swine  erysipelas,  136 
of  swine  plague,  136 
of  symptomatic  anthrax,  133 
of  syphilis,  97 
of  tetanus,  124 
of  typhoid  fever,  101 
phosphorescens  gelidus,  81 

indicus,  80 

indigenus,  81 
pneumo-,  of  Frankel,  112 

of  Friedlander,  111 
potato,  73 
prodigiosus,  72 
pyocvaneus,  120 

0,121 

(155) 


156 


INDEX. 


Bacillus,  ramosus,  74 ' 

root,  74 

smegma,  97 

spinous,  76 

subtilis,  75 

tuberculosis,  88 

violaceus,  79 
Bacteria,  17 

antagonism  of,  67 

asporogenic,  22 

as  remedial  agents,  67 

desmo-,  17 

effect  on  body,  62 

fluorescent,  80 

in  air,  146 

in  milk,  76 

in  water,  79,  149 

infectious,  64 

influence  upon — of  age,  25 

of  electricity,  24 
of  light,  24 
of  oxygen,  24 
of  temperature,  23 

life  of,  23 

micro-,  17 

non-pathogenic,  25,  64 

of  hemorrhagic  septicaemia,  136 

of  pneumonia,  110 

origin  of,  23 

pathogenic,  25,  64 

phosphorescent,  80 

similar  to  cholera  bacillus,  108 

sphere-,  17 

spiro-,  17 

staining  of,  30 

structure  of,  18 

toxic,  64 

unstained,  27 

vital  actions  of,  24 
Bacteridie  du  charbon,  84 
Bacterium  acidi  lactic! ,  77 

aeruginosam,  120 

Balticum,  81 

Fischeri,  81 

Pflugeri,  81 

syncanum,  78 

termo,  139 

ureae,  82 

zopfi,  75 

Beggiatoa  alba,  82 
Benches  for  glass  plates,  56 
Biedert's  method  of  collecting  bac- 
teria, 93 
Black-leg,  133 
Blight,  139 


Blood  serum  as  media,  50 

theory,  65 

Bouillon,  agar,  49 
gelatine,  47 
preparation  of,  44 
sterilization  of,  44 

Bread  mash,  47 

Brood-oven,  50 

Brownian  movements,  19 


CATTLE  plague,  136 
Cell  contents,  18 
Cell  wall,  18 
Cellular  theory,  18,  66 
Charbon  symptomatique,  134 
Charcoal  filter,  150 
Chemical  theory,  65 
Cholera,  105 

red,  108 

Cladothrix  dichotoma,  82 
Classification,  17 
Clostridium,  21 

butyricum,  77 
Cohn's  system,  17 
Cotton  plugs,  43 
Cover-glass  specimens,  34 
Crenothrix,  81 

KUhuiana,  82 
Cultivation,  39 

artificial,  39 

methods  of,  39 

of  anaerobins,  60 
Cultures,  appearances  of,  58 

egg,  52 

filtration  of,  62 

glass  plate,  52 

glass-slide,  52 

potato,  45 

rolled,  57 

test-tube,  52 


DE  BARY'S  system,  18 
Decolorants,  31 
Diphtheria,  101 

Diplococcus  albicans  amplus,  123 
tardissimus,  123 
lanceolatus,  113 
of  pneumonia,  111,  113 
Disease,  cure  of  bacteria,  67 
Disinfectants,  40 

heat  as,  40 
Drying  specimens,  34 


INDEX. 


157 


EUDOSPORUS,  18 
Enteric  fever,  101 
Esmarch's  method,  60 

tubes,  57 
Experiments  on  animals,  68 


FERMENTATION,  25 
I      Fevers,  143 
Filter,  asbestos,  150 

Chamberland,  63 

charcoal,  150 

cotton,  50 

hot  water,  63 

Pasteur,  63 

sand, 148 

sponge,  150 
Filtration  of  cultures,  62 

of  water,  150 
Fishing,  59 
Fission-fungi,  17 
Flagella,  19 

staining  for,  38 
Fluorescence,  25 
Foul-brood,  139 
Frankel's  borer,  153 

method  for  anrerobins,  61 

stain  for  tubercle,  91 
Fuchsin,  carbol,  31,33 
Fungi,  141 
Fungus,  ray,  144 

thrush,  142 


flABBETT'S  stain,  33 
IJ  Gas  formation,  25 
Gelatine,  47 

bouillon,  47 

clouding  of,  48 

paste,  35 

sterilization  of,  49 
Gelatinous  membrane,  18 
Germination,  21 
Gonococcus,  121 
Gonorrhoea,  121 
Gram's  stain,  33 
Guinea-pigs,  68 


H^EMATOMONAS  malarise,  130 
Haematozoa  of  malaria,  130 
Hanging  drop,  29 
Heat,  40 
dry,  40 


leat,  moist,  41 
iemorrhagic  septicaemia,  136 
rlerpes  tonsurans,  144 
ilesse's  method  for  air,  146 

for  anaerobins,  60 
log  cholera,  136 
Homogeneous  lens,  26 
Hot-air  oven,  40 
Hot-water  filter,  63 
Huppe's  method,  61 


IMMERSION  lens,  26 
i.  Immunity,  66 

acquired,  67 

artificial,  67 

natural,  66 
Incubators,  50 
Inoculation,  69 

cutaneous,  69 

in  eye,  70 

intra-duodenal,  70 

intra-peritoneal,  69 

intra-tracheal,  70 

intra-venous,  69 

of  cerebral  membranes,  70 

subcutaneous,  69 
Iodine,  31 
Iris  blender,  27 
Iron  box  for  plates,  55 


KLATSCH  preparations,  59 
Kochin,  95 
Koch's  lymph,  95 
rules,  68 
stain,  32 
steam-chest,  41 
Kiihne's  stain,  33 
method,  38 


LACTIC  acid,  134 
Laveran,  semilunar  bodies  of, 

130 

Leprosy,  96 
Liborius's  method,  60 
Liquefaction  of  media,  25 
Locomotion,  18 
Loffler's  alkaline  stain,  32 
blood  serum,  100 
mordant,  33 
Lupus,  96 
Lustgarten's  method,  97 


158 


INDEX. 


MALARIA,  130 
Material  from  animals,  71 
Media,  fluid,  44 

nutrient,  44 

solid,  45 

transparent,  47 
Metschnikoff's  theory,  66 
Microbe  en  huit,  134 
Micrococci  similar  to  gonococcus, 

123 
Micrococcus  amylivorus,  139 

cerus  albus,  120 
flavus,  120 

cholera  gallinarum,  124 

citreus  conglomeratus,  123 

Indicus,  73 

of  gonorrhoea,  121 

of  mal  de  pis,  138 

of  osteomyelitis,  119 

of  sputum  septicaemia,  111 

Pasteuri,  113 

pyogenes  aureus,  118 
citreus,  120 

subflavus,  124 

tenuis,  120 

tetragenus,  115 

ureae,  82 
Micro-organisms    of    suppuration, 

116 

Microscope,  26 
Microsporon  furfur,  143 
Moist  chamber,  45,  56 
Mordants,  31 
Moulds,  141,  143 

examination  of,  144 
Mouse  septicaemia,  138 
Movements,  vibratory,  19 
Mucor  mucedo,  143 
Mycoprotein,  18 


NAIL  culture,  112 
Nivellier  apparatus,  55 
Nutrient  media,  44 


OIDIUM,  142 
albicans,  14? 
lactis,  142 

Odors  In  cultures,  25 
(Edema  malignans,  127 
Oil  immersion,  26 
Oxidation,  24 


PARASITES,  23 
L     facultative,  'J3 
Pasteur  filter,  63 
Pencillium  glaucum,  143 
Petri's  sand  filter,  148 

saucers,  57 
Phosphorescence,  25 
Pigmentation,  25 
Plasmodium  malaria?,  130 
Platinum  needles,  28 
Pneumo-bacillus,  111 
Potato  cubes,  46 

cultures,  45 

inoculation  of,  46 

in  test-tubes,  46 

mash,  47 

Products  of  tubercular  bacilli,  95 
Proteus,  140 

mirabilis,  140 

vulgaris,  140 

zenkeri,  140 
Ptomaines,  24,  62 
Putrefaction,  25 


RABBIT  septicaemia,  136 
Rausch  brandy,  134 
Reduction,  24 
Relapsing  fever,  129 
Reproduction,  19 
Rouge t  du  pore,  136 


QACCHAROMYCES  albicans,  141 
ij    cerevisia?,  141 

mycoderma,  143 

niger,  141 

rosaceus,  141 
Saprophytes,  23 
Sarcina  alba,  84 

aurantica,  84 

flava,  84 

lutea,  83 

rosea,  84 

ventriculi,  84 
Schizomycetes,  17 
Schweinerotlauf,  136 
Slides,  concave,  29 
Soil,  examination  of,  152 
Solutions,  composite,  31 

formulae  of,  32 

stock,  30,  32 

strong,  32 

weak,  31 


INDEX. 


159 


Soor,  142 
Spasmotoxin,  127 
Specimens,  cover-glass,  34 

cutting  of,  36 

drying  of,  34 

Klatsch,  59 

permanent,  35 
Spirillum,  17 

choleras,  105 

concentricum,  83 

Finkleri,  108 

of  relapsing  fever,  129 

rubrurn,  83 

tyrogenum,  109 
Spirochaete  obermaieri,  129 
Spores,  arthro-,  18,  22 

contents  of,  21 

endo-,  18,  20 

formation  of,  18,  20 

requisites  for,  22 

resistance  of,  22 

staining  of,  37,  38  ' 
Stain,  alkaline,  31,  32 

alkaline  aniline  water,  33 

Gabbet's,  33 

Gram's,  33 

Koch's,  32 

Kuhne's,  33 

Loffler's,  32 

picro-carmine,  33 

Ziehl-Nielsen,  33 
Staining,  Ernst's  method  of,  38 

De  Giacomi's,  98 

general  method  of,  34 

Gram's  method  of,  37 

Kuhne's  method  of,  39 

Loffler's  method  of,  99 

Lustgarten's  method  of,  97 

of  flagella,  38 

of  spores,  37,  38 

of  sporogenic  bodies,  38 

of  tissue  sections,  36 

rapid  method  for  tubercle,  92 

slow  method  of,  for  tubercle,  93 

solutions,  30 

special  methods  of,  36 

Weigert's  method  of,  38 
Staphylococcus,  18 

pyogenes  aureus,  118 


Sterilization,  39 

fractional,  42 
Streptococci,  18 

in  diphtheria,  101 
Streptococcus  pyogenes,  117 
Suppuration,  116 
Swine  diphtheria,  136 

erysipelas,  136 

plague,  136 
Syphilis,  97 
System  of  Cohn,  17 

of  De  Bary,  18 


rpEST-TUBES,  43 
1     Tetanin,  127 
Tetanis,  124 
Tetanotoxin,  127 
Thermo-regulator,  51 
Thrush,  142 
Tinea,  144 

Toxalbumens,  62,  67 
Toxines,  62 

Tricophyton  tonsurans,  143 
Tuberculin,  67,  95 
Tuberculosis,  88 
Typhoid  fever,  101 
Typhotoxine,  104 


VIBRIO,  Finkler-Prior,  108 
Metschnikoff,  110 
Vibrion  septique,  127 


WATER,  bacteria  in,  77,  152 
examination  of,  149 
Weigert's  method  of  staining,  38 
Wire  cages,  43 
Wolfhugel's  apparatus,  152 


YEASTS,  141 
JL     examination  of,  144 


F/IEHL'S  solution,  33 
Zj    Zooloea,  18 


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2 


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and  Dissector— this  is  a  good  dissector's  manual,  with  clear  type  and  hand- 
some cuts.  The  colored  plates  are  especially  commendable. 
•  The  Southern  Practitioner,  Nashville,  Tenn.,  September,  1890.— Nancrede's 
Anatomy  and  Dissector— truly  a  "  Vade  Mecum,"  a  "  multum  in  parvo."  The 
illustrations  are  marvels  of  beauty  and  clearness  of  illustration. 

7 


IN  PREPARATION. 


DISEASES  OF  THE  EYE. 

BY 

G.  E.  DE  SCHWEINITZ,  M.D., 

Ophthalmic  Surgeon  to  Children's  Hospital  and  to  the  Philadelphia  Hospital 
Ophthalmologist  to  the  Orthopedic  Hospital  and  Infirmary  for  Ner- 
vous Diseases;   Lecturer  on  Medical   Ophthalinoscopy, 
University  of  Pennsylvania,  etc. 


A  HAND-BOOK  OF  OPHTHALMIC  PRACTICE, 

Especially  useful  to  the  student  who  has  Imd  neither  time 
nor  inclination  to  study  the  numerous  able  but  more  volu- 
minous text-books. 


The  object  of  this  manual  is  to  present  to  the  student  who  is  be- 
ginning work  in  the  field  of  ophthalmology  a  plain  description  of 
the  optical  defects  and  diseases  of  the  eye.  To  this  end  special 
attention  has  been  paid  to  the  clinical  side  of  the  question  ;  and  the 
methods  of  examination,  the  symptomatology  leading  to  a  diagnosis, 
and  the  treatment  of  the  various  ocular  defects  have  been  brought 
into  special  prominence.  Anatomy,  physiology,  and  pathological 
histology,  except  in  so  far  as  they  serve  the  purpose  just  stated, 
have  been  omitted.  The  sections  devoted  to  optical  principles  and 
the  normal  and  abnormal  refraction  of  the  eye  in  large  portion  have 
betn  written  by  Dr.  James  Wallace,  Chief  of  the  Eye  Dispensary  of 
the  University  Hospital.  The  chapter  devoted  to  the  application 
of  the  shadow-test  has  been  prepared  by  Dr.  Edward  Jackson.  The 
book  will  be  suitably  illustrated  by  a  number  of  wood-cuts,  many  of 
them  from  cases  in  the  practice  of  the  author,  in  addition  to  which 
there  will  be  several  chromo-lithographs. 


IN  PREPARATION. 


DISEASES  OF  WOMEN. 

BY  HENRY  J.  GARRIGUES,  A.M.,  M.D., 

Professor  of  Obstetrics  in  the  New  York  Post-Graduate  Medical  School  and 
Hospital ;  Gynaecologist  to  St.  Mark's  Hospital  in  New  York  City  ;  Gynae- 
cologist to  the  German  Dispensary  in  the  City  of  New  York;  Con- 
sulting Obstetrician  to  the  New  York  Infant  Asylum;  Obstetric 
Surgeon  to  the  New  York  Maternity  Hospital ;  Fellow  of 
the  American  Gynaecological  Society  ;  Fellow  of  the 
New  York  Academy  of  Medicine  ;  President  of  the 
German  Medical  Society  of  the  City  of  New 
York,  etc.  etc. 

It  is  the  intention  of  the  writer  to  provide  a  practical  manual  on 
Gynaecology,  for  the  use  of  students  and  practitioners,  in  as  concise  a 
manner  as  is  compatible  with  clearness. 


Syllabus  of  Obstetrical  Lectures 

In  the  Medical  Department,  University  of  Pennsylvania, 

BY  RICHARD  C.  NORRIS,  A.M.,  M.D., 

DEMONSTRATOR  ON  OBSTETRICS  IN  THE  UNIVERSITY  OF  PENNSYLVANIA. 


Price,  Cloth,  Interleaved  for  Notes  .  ,  ,  $2.00  Net, 

The  New  York  Medical  Record  of  April  19,  1890,  referring  to  this 
book,  says  :  "  This  modest  little  work  is  so  far  superior  to  others  on 
the  same  subject  that  we  take  pleasure  in  calling  attention  briefly  to 
its  excellent  features.  Small  as  it  is,  it  covers  the  subject  thoroughly, 
and  will  prove  invaluable  to  both  the  student  and  the  practitioner  as 
a  means  of  fixing  in  a  clear  and  concise  form  the  knowledge  derived 
from  a  perusal  of  the  larger  text-books.  The  author  deserves  great 
credit  for  the  manner  in  which  he  has  performed  his  work.  He  has 
introduced  a  number  of  valuable  hints  which  would  only  occur  to  one 
who  was  himself  an  experienced  teacher  of  obstetrics.  The  subject- 
matter  is  clear,  forcible,  and  modern.  We  are  especially  pleased  with 
the  portion  devoted  to  the  practical  duties  of  the  accoucheur,  care  of 
the  child,  etc.  The  paragraphs  on  antiseptics  are  admirable  ;  there 
is  no  doubtful  tone  in  the  directions  given.  No  details  are  regarded 
as  unimportant  ;  no  minor  matters  omitted.  We  venture  to  say  that 
even  the  old  practitioner  will  find  useful  hints  in  this  direction  which 
he  cannot  afford  to  depise." 

9 


READY  SHORTLY. 


SAUNDERS' 

Pocket  Medical  Formulary. 

BY 

WILLIAM  M.  POWELL,  M.D., 

Attending  Physician  to  the  Mercer  House  for  Invalid  Women,  at  Atlantic 

City,  N.  J.  ;  Late  Physician  to  the  Clinic  for  the  Diseases  of  Children 

in  the  Hospital  of  the  University  of  Pennsylvania  and  St.  Clement's 

Hospital ;  Instructor  in  Physical  Diagnosis  in  the  Medical 

Department  of  the  University  of   Pennsylvania,  and 

Chief  of  the  Medical  Clinic  of  the  Philadelphia 

Polyclinic. 


Containing  about  2000  Formulae,  selected  from  several 
hundreds  of  the  best-known  authorities. 

A  concise,  clear,  and  correct  record  of  the  many 
hundreds  of  famous  formulae  which  are  found  scattered 
through  th°!  works  of  the 

Most  imminent  Physicians  and  Surgeons 

of  the  world ;  particularly  helpful  to  the  student  and 
young  practitioner,  as  it  gives  him  a  taste  for  writing  his 
prescriptions  in  an  elegant  and  correct  manner,  thus  avoid- 
ing incompatible  and  dangerous  prescriptions.  The  use 
of  this  work  is  to  be  recommended  even  to  the  older  prac- 
tioner,  as  through  it  he  becomes  acquainted  with  numerous 
formulae  which  are  not  found  in  the  text-books,  but  have 
been  collected  from  among  the 

Rising  Generation  of  the  Profession,  College  Professors,  and 

Hospital  Physicians  and  Surgeons. 

10 


NOW  READY. 


NEW  AND  REVISED  EDITIONS  OF 

SAUNDERS' 

QUESTION  COMPENDS, 

Arranged  in  Question  and  Answer  Form. 

The  Latest,  Cheapest,  and  Best 
ILLUSTRATED  SERIES  OF  COMPENDS  EVER  ISSUED. 


THE  ADVANTAGES  OF  QUESTIONS  AND 
ANSWERS. — The  usefulness  of  arranging  the  subjects  in 
the  form  of  Questions  and  Answers  will  be  apparent, 
since  the  student,  in  reading  the  standard  works,  often  is  at 
a  loss  to  discover  the  important  points  to  be  remembered, 
and  is  equally  puzzled  when  he  attempts  to  formulate  ideas 
as  to  the  manner  in  which  the  Questions  could  be  put 
in  the  Examination-Room. 


These  small  works,  which  can  be  conveniently  carried  in  the  pocket, 
contain  in  a  condensed  form  the  teachings  of  the  most  popular 
text  books. 

The  authors  are  nearly  all  connected  with  the  various  colleges  as 
Demonstrators  or  Lecturers,  and  are  therefore  thoroughly  conver- 
sant, not  only  with  the  wants  of  the  average  student,  but  also  with 
the  points  that  are  absolutely  necessary  to  be  remembered  in 
the  Examination-Room.  These  books  are  constantly  in  the  hands 
of  their  authors  for  revision,  and  are  kept  well  up  to  the  times,  their 
fast  sale  allowing  them  to  be  almost  entirely  rewritten  whenever 
necessary,  instead  of  having  to  wait  for  the  edition  to  be  sold,  as  is 
the  case  with  an  ordinary  text-book. 

11 


No.  1. 

ESSEITIALS  OF  PHYSIOLOGY, 


H.  A.  HARE,  M.D., 

Professor  of  Therapeutics  and  Materica  Medica  in  the  Jefferson  Medical  Col. 

lege  of  Philadelphia;  Physician  to  St.  Agnes'  Hospital  and  to  the 

Medical  Dispensary  of  the  Children's  Hospital ;  Laureate  of 

the  Royal  Academy  of  Medicine  in  Belgium,  of  the 

Medical  Society  of  London,  etc. ;  Secretary 

of  the  Convention  for  the  Revision  of 

the  Pharmacopoeia,  1S90. 

NUMEROUS  ILLUSTRATIONS. 

Third  Edition,  Revised  and  Enlarged. 

Price,  Cloth    .         .    $1.00;   Interleaved  for  Notes   .         .    $1.25. 


Unii-ersity  Medical  Magazine, 
October,  1888.— "  Dr.  Hare  has 
admirably  succeeded  in  gather- 
ing together  a  series  of  Ques- 
tions which  are  clearly  put  and 
tersely  answered." 

Pacific  Medical  Jmirnal,  Octo- 
ber, 1889.—"  Hare's  Physiology 
contains  the  essences  of  its  sub- 
ject. No  better  book  lias  ever 
been  produced,  and  every  stu- 
dent would  do  well  to  possess  a 
copy." 

Times  and  Register,  Philadel- 
phia, October  ft,  1889.—"  In  the 
second  edition  of  Hare's  Physi- 
ology all  the  moredifficult  points 
of  the  study  of  the  nervous  sys- 
tem have  been  elucidated.  As 
the  work  now  appears  it  cannot 
fail  to  merit  the  appreciation  of 

Specimen  of  Illustrations.  the  overworked  student." 

Journal  of  the  American  Association,  November  23,  1889. — "  Hare's 
Physiology — an  excellent  work  ;  admirably  illustrated  ;  well  calcu- 
lated to  lighten  the  task  of  the  over-burdened  undergraduate." 

12 


No.  2. 

ESSENTIALS  OF  SURGERY. 

CONTAINING,  ALSO, 

Venereal  Diseases,  Surgical  Landmarks,  Minor  and  Operative  Su~- 
gery,  and  a  Complete  Description,  together  with  full  Illustra- 
tions, of  the  Handkerchief  and  Roller  Bandage. 

BY  EDWARD  MARTIN,  A.M.,  M.D., 

Clinical  Professor  of  Genito-Uriiiary  Diseases,  Instructor  in  Operative  Sur- 
gery, and  Lecturer  on  Minor  Surgery,  University  of  Pennsylvania; 
Surgeon  to  the  Howard  Hospital ;  Assistant  Surgeon  to  the 
University  Hospital,  etc.  etc. 


PROFUSELY   ILLUSTRATED. 

FOURTH  EDITION, 

Considerably  enlarged  by  an  Appendix  containing  Ml  directions 
and  prescriptions  for  the  preparation  of  the  various  mate- 
rials used  in  ANTISEPTIC  SURGERY  ;  also  sev- 
eral hundred  recipes  covering  the  medical 

treatment  of  surgical  affections. 
Price,  Cloth,  $1.00.     Interleaved  for  Notes,  $1.25. 

Me.lical  and  Surgical  Rejjorter, 
February,  1889.  —  "  Martin's  Sur- 
gery contains  all  necessary  essen- 
tials of  modern  surgery  in  a  com- 
paratively small  space.  Its  style 
is  interesting  and  its  illustrations 
admirable." 

University  Medical  Magazine, 
January,  1889.— "Dr.  Martin  has 
admirably  succeeded  in  selecting 
and  retaining  just  what  is  neces- 
sary for  purposes  of  examination, 
and  putting  it  in  most  excellent 
shape  for  reference  and  memor- 
izing." 

Kansas  City  Medical  Record. — 
"Martin's  Surgery. — This  admir- 
able compend  is  well  up  in  the 
most  advanced  ideas  of  modern 

surgery."  Specimen  of  Illustrations. 

13 


No.  3. 

ESSENTIALS  OF  ANATOMY, 

Including  the  Anatomy  of  the  Viscera. 

BY  CHARLES  B.  NANCREDE,  M.D., 

Professor  of  Surgery  and  Clinical  Surgery  in  the  University  of  Michigan, 

Ann  Arbor ;  Corresponding  Member  of  the  Royal  Academy  of 

Medicine,  Rome,  Italy ;  Late  Surgeon  Jefferson 

Medical  College,  etc.  etc. 

ONE  HUNDRED  AND  FORTY  FINE  WOODCUTS 

THIRD  EDITION. 
Enlarged  by  an  Appendix  containing  over  Sixty  Illustrations  of 

the  Osteology  of  the  Human  Body. 
The  whole  based  upon  the  last  (eleventh)  edition  of 

GRAY'S  ANATOMY. 
Price,  Cloth,  $1.00.    Interleaved  for  Notes,  $1.25. 

American  Practitioner   and 
News,  February  16,  18b9. 

"  Nancrede's  Anatomy. — 
For  self-quizzing  and  keep- 
ing fresh  in  mind  the 
knowledge  of  Anatomy 
gains  at  school,  it  would 
not  be  easy  to  speak  of  it 
in  terms  too  favorable." 

Southern   Californian   Practi- 
tioner, January  18,  1880. 
"  Nancrede's  Anatomy. — 
Very   accurate   and    trust- 
worthy." 

American  Practitioner  and 
Neu:s,  Louisville,  Kentucky. 
"  Nancrede's  Anatomy. — 
Truly  such  a  book  as  no 
student  can  afford  to  be 
without." 


Specimen  of  Illustrations. 


14 


No.  4. 


Essentials  of  Medical  Chemistry 

ORGANIC  AND  INORGANIC. 


CONTAINING,  ALSO, 

Questions  on  Medical  Physics,  Chemical  Physiology, 
Analytical  Processes,  Urinalysis,  and  Toxicology. 

BY 

LAWRENCE  WOLFF,  M.D., 

Demonstrator  of  Chemistry,  Jefferson  Medical  College  ;  Visiting  Physician 

to  German  Hospital  of  Philadelphia  ;  Member  of  Philadelphia 

College  of  Pharmacy,  etc.  etc. 

SIXTH  THOUSAND. 


Price,  Cloth,  $1.00.     Interleaved  for  Notes,  $1.25. 


Cincinnati  Medical  News,  January,  1889.  —  "  Wolff's  Chemistry.-  -A  little 
work  that  can  be  carried  in  the  pocket,  for  ready  reference  in  solving  difficult 
problems." 

St..  Joseph's  Medical  Herald,  March,  1889.— "Dr.  Wolff  explains  most 
simply  the  knotty  and  difficult  points  in  chemistry,  and  the  book  is  therefore 
well  suited  for  use  in  medical  schools." 

Medical  and  Surgical  Reporter,  November,  1889. — "We  could  wish  that 
more  books  like  this  would  be  written,  in  order  that  medical  students  might 
thus  early  become  more  interested  in  what  is  often  a  difficult  and  uninterest- 
ing branch  of  medical  study." 

Registered  Pharmacist,  Chicago,  December,  1890.— "  Wolff 's  Chemistry." 
— "  The  author  is  thoroughly  familiar  with  his  subjects.  A  useful  addition  to 
the  medical  and  pharmaceutical  library." 

15 


No.  5. 

ESSENTIALS  OE  OBSTETRICS, 

BY  W.  EASTERLY  ASHTON,  M.D., 

Obstetrician  to  the  Philadelphia  Hospital 


NUMEROUS  ILLUSTRATIONS.     SIXTH  THOUSAND. 


Price,  Cloth,  $1.00.     Interleaved  for  Notes,  $1.25. 

A 


Specimen  of  Illustrations. 

Southern  Practitioner,  January,  1890. — Ashton's  Obstetrics. — An  excellent 
little  volume  containing  correct  and  practical  knowledge.  An  admirable  com- 
pend,  and  the  best  condensation  we  have  seen." 

Chicago  Medical  Times. — "  Ashton's  Obstetrics. — Of  extreme  value  to  stu- 
dents, and  an  excellent  little  book  to  freshen  up  the  memory  of  the  practi- 
tioner." 

Medical  and  Surgical  Reporter,  January  26,  1889. — "  Ashton's  Obstetrics. 
— A  work  thoroughly  calculated  to  be  of  service  to  students  in  preparing  for 
examination.7* 

.\nr  York  M&liral  Abstract,  April,  1890.— «« Ashton's  Obstetrics  should  be 
consulted  by  the  medical  student  until  he  can  answer  every  question  at  sight. 
The  practitioner  would  also  do  well  tb  glance  at  the  book  now  and  then,  to 
prevent  his  knowledge  from  getting  rusty." 

16 


No.  6. 

ESSENTIALS 

OP 

Pathology  and  Morbid  Anatomy. 


C.  E.  A  It  JI A  XI)  SEMl'LE,  B.A,  M.B.,  Cantab.,  L.S.A.,  M.R.C.P.,  Innil., 

Physician  to  the  Northeastern  Hospital  for  Children,  Harkney  ;   Pro- 
fessor of  Vocal  and  Aural  Physiology  and  Examiner  in  Acous- 
tics at  Trinity  College,  London,  etc.  etc. 


ILLUSTRATED.    FOURTH  THOUSAND, 


Price,  Cloth,  $1.OO.    Interleaved  for  Notes,  $1.25. 

From  the  College  and  (Jlimcul  Record, 
September,  1889. — "  A  small  work  upon 
Pathology  and  Morbid  Anatomy,  that  re- 
duces such  complex  subjects  to  the  ready 
comprehension  of  the  student  and  practi- 
tioner, is  a  very  acceptable  addition  to 
medical  literature.  All  the  more  modern 
topics,  such  as  Bacteria  and  Bacilli,  and 
the  most  recent  views  as  to  Urinary  Path- 
ology, find  a  place  here,  and  in  the  hands 
of  a  writer  and  teacher  skilled  in  the  art 
of  simplifying  abstruse  and  difficult  sub- 
jects for  easy  comprehension  are  rendered 
thoroughly  intelligible.  Few  physicians 
do  more  than  refer  to  the  more  elaborate 
works  for  passing  information  at  the  time 
it  is  absolutely  needed,  but  a  book  like  this 
of  Dr.  Semple's  can  be  taken  up  and  perused  continuously  to  the  profit  and 
instruction  of  the  reader." 

Indiana,  Medical  Journal,  December.  1889. — "Semple's  Pathology  and 
Morbid  Anatomy. — An  excellent  compend  of  the  subject  from  the  points  of 
view  of  Green  and  Payne." 

Cincinnati  Medical  News,  November,  1889. — Semple's  Pathology  and  Mor- 
bid Anatomy. — A  valuable  little  volume — truly  a  muUnm.  in  paivo." 

17 


Specimen  of  Illustrations. 


No.  7. 

ESSENTIALS 

OF 

Materia  Medica,  Therapeutics, 

AND 

PRESCRIPTION  WRITING. 


HENRY  MORRIS,  M.D., 

Late  Demonstrator,  Jefferson  Medical  College  ;  Fellow  College  of  Physicians, 

Philadelphia;  Co-editor  Biddle's  Materia  Medica ;  Visiting 

Physician  to  St.  Joseph's  Hospital,  etc.  etc. 


SECOND  EDITION.     FOURTH  THOUSAND. 


Price,  Cloth,  $1.00.     Interleaved  for  Notes,  $1.25. 


MEDICAL  AND  SURGICAL  REPORTER,  October,  1889. 

"Morris*  Materia  Medica  and  Therapeutics. —One  of  the  best  compends  in 
this  series.  Concise,  pithy,  and  clear,  well-suited  to  the  purpose  for  which  it 
is  prepared." 

GAILLARD'S  MEDICAL  JOURNAL,  November,  1889. 

"  Morris*  Materia  Medica.— The  very  essence  of  Materia  Medica  and  Thera- 
peutics boiled  down  and  presented  in  a  clear  and  readable  style." 

SANITARIUM,  New  York,  January,  1890. 

"Morris*  Materia  Medica.— A  well-arranged  quiz-book,  comprising  the 
most  important  recent  remedies." 

BUFFALO  MEDICAL  AND  SURGICAL  JOURNAL,  January,  1890. 
"Morns'  Materia  Medica. — The  subjects  are  treated  in  such  a  unique  and 
attractive  manner  that  they  cannot  fail  to  impress  the  mind  and  instruct  in 
a  lasting  manner." 

18 


Nos.  8  and  9. 

Essentials  of  Practice  of  Medicine, 

BY  HENRY  MORRIS,  M.D., 

Author  of  "  Essentials  of  Materia  Medica,"  etc. 

With  an  Appendix  on  the  Clinical  and  Microscopical 
Examination  of  Urine. 

BY  LAWRENCE  WOLFF,  M.D., 

Author  of  "Essentials  of  Medical  Chemistry,"  etc. 


COLORED  (VOGEL)  URINE  SCALE  AND  NUMEROUS 
FINE  ILLUSTRATIONS. 


8ECOND    EDITION, 

Enlarged  by  some  THREE   HUNDRED  Essential 

Formulae,  selected  from  the  writings  of  the 

most  eminent  authorities  of  the 

Medical  Profession. 

COLLECTED  AND  ARRANGED  BY 

WILLIAM  M.  POWELL,  M.D., 

Author  of  "Essentials  of  Diseases  of  Children.'' 


Price,  Cloth,  &2.OO.    Medical  Sheep,  $*2.5O. 

SOUTHERN  PRACTITIONER,  Nashville,  Tenn.,  January,  1891. 
"Morris*  Practice  of  Medicine. — Of  material  aid  to  the  advanced  student 
in  preparing  for  his  degree,  and  to  the  young  practitioner  in  diagnosing  affec- 
tions or  selecting  the  proper  remedy." 

AMERICAN  PRACTITIONER  AND  NEWS,  Louisville,  Ky.f  January,  1891. 
"Morris'  Practice  of  Medicine. — The  teaching  is  sound,  the  presentation 
graphic,  matter  as  full  as  might  be  desired,  and  the  style  attractive." 

SOUTHERN  MEDICAL  RECORD,  January,  1891. 

"Morris*  Practice  of  Medicine  is  presented  to  the  reader  in  the  form  of 
Questions  and  Answers,  thereby  calling  attention  to  the  most  important  lead- 
ing facts,  which  is  not  only  desirable,  but  indispensable  to  an  acquaintance 
with  the  essentials  of  medicine.  The  book  is  all  it  pretends  to  be,  and  we 
cheerfully  recommend  it  to  medical  students." 

19 


No.  10. 

ESSENTIALS  OF  GYNAECOLOGY, 


EDWIN  B.  CRAIGIN,  M.D., 

Attending  Gynaecologist,    Roosevelt    Hospital,    Out- Patients'    Department 
Assistant  Surgeon,  New  York  Cancer  Hospital,  etc.  etc. 

58  FINE  ILLUSTRATIONS. 
SIXTH  THOUSAND. 


Price,  Cloth,  $1.OO.    Interleaved  for  Notes,  $1.25. 


m 


•m* 


Specimen  of  Illustrations. 


Medical  and  Surgical  Re- 
porter, April, 1890.— "Craig- 
gin's  Essentials  of  Gynaecol- 
ogy.— This  is  a  most  excel- 
lent addition  to  this  series 
of  question  compends,  and 
properly  used  will  be  of 
great  assistance  to  the  stu- 
dent in  preparing  for  ex- 
amination. Dr.  Craigin  is 
to  be  congratulated  upon 
having  produced  in  com- 
pact form  the  Essentials  of 
Gynaecology.  The  style  is 
concise,  and  at  the  same 
time  the  sentences  are  well 
rounded.  This  renders  the 
book  far  more  easy  to  read 
than  most  compends,  and 
adds  distinctly  to  its  value." 

College  and  Clinical  Record, 
April,  1890.  —  "  Craigin's 
Gynaecology. — Students  and 
practitioners,  general  or  spe- 
cial, even  derive  information 
and  benefit  from  the  perusal 
and  study  of  a  carefully 
written  work  like  this." 


No.  11. 

Essentials  of  Diseases  of  the  Skin, 

BY  HENRY  W.  STELWAGON,  M.D., 

Clinical  Lecturer  on  Dermatology  in  the  Jefferson  Medical  College,  Philadel- 
phia; Physician  to  Philadelphia  Dispensary  for  Skin  Diseases;  Chief 
of  the  Skin  Dispensary  in  the  Hospital  of  University  of  Penn- 
sylvania; Physician  to  Skin  Department  of  the  Howard 
Hospital ;  Lecturer  on  Dermatology  in  the  Women's 
Medical  College,  Philadelphia,  etc.  etc. 

74  ILLUSTRATIONS,  many  of  which  are  original. 


FOURTH  THOUSAND. 


Price,  Cloth,  $1.00.    Interleaved  for  Notes,  $1.25. 


Specimen  of  Illustrations. 

New  York  Medical  Journal,  May,  1890.— "Stelwagon's  Diseases  of  the 
Skin.— We  are  indebted  to  Philadelphia  for  another  excellent  book  on  Derma- 
tology. The  little  book  now  before  us  is  well  entitled  "Essentials  of  Derma- 
tology," and  admirably  answers  the  purpose  for  which  it  is  written."  The 
experience  of  the  reviewer  has  taught  him  that  just  such  a  book  is  needed. 
We  are  pleased  with  the  handsome  appearance  of  the  book,  with  its  clear 
type,  good  paper,  and  fine  wood-cuts." 


No.  12. 

ESSENTIALS 


Minor  Surgery,  Bandaging,  and 
Venereal  Diseases, 

BY  EDWARD  MARTIN,  A.M.,  M.D., 

Author  of  "Essentials  of  Surgery,"  etc. 

82  ILLUSTRATIONS,  mostly  specially  prepared  for  this  wcrk. 


Price,  Cloth,  $1.00.     Interleaved  for  Notes,  $1.25. 


Medical    News,    Phila- 
delphia, January  10,1891. 
"Martin's  Minor  Surgery, 
Bandaging,  and  Venereal 
Diseases. —  The  best  con- 
densation of  the  subjects 
of  which  it  treatsyetplaced 
before  the  profession.    The 
chapteronGenito-Urinary 
Diseases,  though  short,  is 
sufficiently     complete     to 
'  ~Y"\  make    them     thoroughly 
I  acquainted  with  the  most 
_  /   /  advanced    views    on    the 
subject." 

Nashville    Journal    of 
Medicineaiid  Surgery,  No- 
vember, 1 890. — '  'Martin 's 
Minor  Surgery, etc., should 
be  in  the  hands  of  every 
student,  and  we  shall  per- 
sonally recommend  it  toour 
students  as  the  best  text- 
book upon  the  subject." 
Pharmaceutical  Era,  Detroit,  Michigan,  December  1,  1890. — "Martin's 
Minor  Surgery,  etc. — Especially  acceptable  to  the  general  practitioner,  who 
is  often  at  a  loss  in  cases  of  emergency  as  to  the  proper  method  of  applying  a 
bandage  to  an  injured  member." 

22 


Specimen  of  Illustrations. 


No.  13. 

ESSENTIALS 


OJT 

Legal  Medicine,  Toxicology, 


HYGIENE. 

BY 

C.  E.  ARMAND  SEMPLE,  M.D., 

Author  of  "  Essentials  of  Pathology  and  Morbid  Anatomy." 


130  ILLUSTRATIONS. 


Price,  Cloth $1.OO. 

Interleaved  for  Notes         ....       1.25. 


SOUTHERN  PRACTITIONER,  Nashville,  May,  1890. 
"Seraple's  Legal  Medicine,  etc. — At  the  present  time,  when  the 
field  of  medical  science,  by  reason  of  rapid  progress,  becomes  so  vast, 
a  book  which  contains  the  essentials  of  any  branch  or  department  of 
it,  in  concise,  yet  readable  form,  must  of  necessity  be  of  value.  This 
little  brochure,  as  its  title  indicates,  covers  a  portion  of  medical  science 
that  is  to  a  great  extent  too  much  neglected  by  the  student,  by  reason 
of  the  vastness  of  the  entire  field  and  the  voluminous  amount  of  matter 
pertaining  to  what  he  deems  more  important  departments.  The  lead- 
ing points,  the  essentials,  are  here  summed  up  systematically  and 
clearly." 

MEDICAL  BRIEF,  St.  Louis,  May,  1890. 

"  Semple's  Legal  Medicine,  Toxicology,  and  Hygiene. — A  fair  sample 
of  Saunders'  valuable  compends  for  the  student  and  practitioner.  It 
is  handsomely  printed  and  illustrated,  and  concise  and  clear  in  its 

teachings." 

23 


No.  14. 

ESSENTIALS  OF 

Refraction  and  Diseases  of  the  Eye, 

BY  EDWARD  JACKSON,  A.M.,  M.D., 

Professor  of  Diseases  of  the  Eye  in  the  Philadelphia  Polyclinic  and  College  for 
Graduates  in  Medicine ;  Member  of  the  American  Ophthalmological  So- 
ciety; Fellow  of  the  College  of  Physicians  of  Philadelphia;  Fel- 
low of  the  American  Academy  of  Medicine,  etc.  etc. 

AND 

Essentials  of  Diseases  of  the  Nose  and  Throat, 

BY  E.  BALDWIN  GLEASON,  M.D., 

Assistant  in  the  Nose  and  Throat  Dispensary  of  the  Hospital  of  the  University 

of  Pennsylvania ;  Assistant  in  the  Nose  and  Throat  Department  of  the 

Union  Dispensary;  Member  of  the  German  Medical  Society, 

Philadelphia  ;  Polyclinic  Medical  Society,  etc.  etc. 

TWO  VOLUMES  IN  ONE.     PROFUSELY  ILLUSTRATED. 


Price,  Cloth,  91.OO.     Interleaved   for   Notes,  $1.25. 


University  Medical  Mag- 
azine, Philadelphia,  Octo- 
ber, 1890. — "Jackson  and 
Gleason's  Essentials  of  Dis- 
eases of  the  Eye,  Nose,  and 
Throat.  —  The  subjects 
have  been  handled  with 
skill,  and  the  student  who 
acquires  all  that  here  lays 
before  him  will  have  much 
more  than  a  foundation  for 
future  work." 

New  York  Medical  Rec- 
ord, November  15,  1890. 
— "Jackson  and  Gleason 
on  Diseases  of  the  Eye, 
Nose,  and  Throat.  —  A 

valuable   book  to  the  be- 
Specimen  of  Eye  Illustrations. 

ginner  in  these  branches, 

to  the  student,  to  the  busy  practitioner,  and  as  an  adjunct  to  more  thorough 
reading.  The  authors  are  capable  men,  and  as  successful  teachers  know 
what  a  student  most  needs" 

114 


No.  15. 

ESSENTIALS 


DISEASES  OF  CHILDREN, 


BY 

WILLIAM  M.  POWELL,  M.D., 

Attending  Physician  to  the  Mercer  House  for  Invalid  Women,  at  Atlantic 
City,  N.  J.  ;  Late  Physician  to  the  Clinic  for  the  Diseases  of  Chil- 
dren in  the  Hospital  of  the  University  of  Pennsylvania  and 
St.  Clement's  Hospital ;  Instructor  in  Physical  Diag- 
nosis in  the  Medical  Department  of  the  Uni- 
versity of  Pennsylvania,  and  Chief  of 
the  Medical  Clinic  of  the  Phil- 
adelphia Polyclinic. 


Price,  Cloth        .        .        .       .       .       .        $1.OO. 

Interleaved  for  Notes    ....          1.25. 


AMERICAN  PRACTITIONER  AND  NEWS,  Louisville,  Ky.,  December  20, 1890. 
"  Powell's  Diseases  of  Children. — This  work  is  gotten  up  in  the 
clear  and  attractive  style  that  characterizes  the  Saunders'  Series.  It 
contains  in  appropriate  form  the  gist  of  all  the  best  works  in  the  de- 
partment to  which  it  relates." 

SOUTHERN  PRACTITIONER,  Nashville,  Tennessee,  November,  1890. 
"Dr.  Powell's  little  book  is  a  marvel  of  condensation.     Handsome 
binding,  good  paper,  and  clear  type  add  to  its  attractiveness." 

ANNALS  OF  GYN^COLOGY,  Philadelphia,  December,  1890. 
14  Powell's  Diseases  of  Children. — The  book  contains  a  series  of  im- 
portant questions  and  answers,  which  the  student  will  find  of  great 
utility  in  the  examination  of  children." 


No.  16. 

ESSENTIALS 


EXAMINATION  OF  TJEIIE. 


BY 


LAWRENCE  WOLFF,  M.D., 

Author  of  "Essentials  of  Medical  Chemistry,"  etc. 


COLORED  (VOGEL)  URINE   SCALE  AND  NUMEROUS 
ILLUSTRATIONS. 


Price,  Cloth 


75  Cents. 


Specimen  of  Illustrations. 


UNIVERSITY  MEDICAL  MAGAZINE, 

June,  1890. 

**  Wolff 's  Examination  of  the 
Urine. — A  little  work  of  decided 
value." 

MEDICAL   RECORD,   New  York, 

August  23,  1890. 
"Wolff's  Examination  of 
Urine.  —  A  good  manual  for 
students,  well  written,  and 
answers,  categorically,  many- 
questions  beginners  are  sure 
to  ask." 


MEMPHIS  MEDICAL  MONTHLY,  Memphis,  Tennessee,  June,  1890. 
"Wolff's  Examination  of  Urine. — The  book  is  practical  in  char- 
acUr,  comprehensive  as  is  desirable,  and  a  useful  aid  to  the  student 

in  his  studies." 

20 


No.  18. 

ESSENTIALS 


PRACTICE  OF  PHARMACY, 


BY 


LUCIUS   E.  SAYRE, 

Professor  of  Pharmacy  and  Materia  Medica  in  the  University  of  Kansas. 


Price,  Oloth,  $1,00.    Interleaved  for  Notes,  $1.25, 


ALBANY  MEDICAL  ANNALS,  Albany,  N.  Y.,  November,  1890. 
"  Sayre's  Essentials  of  Pharmacy  covers  a  great  deal  of  ground  in 
small   compass.     The   matter  is  well  digested  and  arranged.     The 
research  questions  are  a  valuable  feature  of  the  book." 

AMERICAN  DOCTOR,  Richmond,  Va.,  January,  1891. 
"  Sayre's  Essentials  of  Pharmacy. — This  very  valuable  little  manual 
covers  the  ground  in  a  most  admirable  manner.     It  contains  practical 
pharmacy  in  a  nutshell." 

NATIONAL  DRUG  REGISTER.  St.  Louis,  Mo.,  December  1,  1890. 
"  Sayre's  Essentials  of  Pharmacy.— The  best  quiz  on  pharmacy  we 
have  yet  examined." 

WESTERN  DRUG  RECORD,  November  10,  1890. 

"Sayre's  Essentials  of  Pharmacy. — A  book  of  only  180  pages,  but 
pharmacy  in  a  nut-shell.  It  is  not  a  quiz-compend  compiled  to  en- 
able a  grocery  clerk  to  '  down'  a  board  of  pharmacy ;  it  is  a  finger- 
post guiding  a  student  to  a  completer  knowledge." 

27 


SAUNUERS'  QUESTION-COMPENDS. 

In  Preparation.    Ready  about  September  1,  1891. 


No.  17. 


Essentials  of  Diagnosis, 


No.  19 


Essentials  of  Hygiene. 

ILLUSTRATED. 

BY  ROBERT  P.  ROBINS,  M.D. 


No.  20. 

Essentials  of  Bacteriology. 

ILLUSTRATED. 

BY  M.  V.  BALL,  M.D. 


No.  21. 

Essentials  of  Nervous  Diseases  and  Insanity, 

ILLUSTRATED. 
BY  JOHN  C.  SHAW,  M.D. 


No.  22. 

Essentials  of  Medical  Physics. 

ILLUSTRATED. 
BY  FRED.  J    BROCKWAY,  M.D. 


No.  23. 


Essentials  of  Medical  Electricity. 

ILLUSTRATED. 

BY  DAVID  D.  STEWART,  M.D.,  and  EDWARD  S.  LAWRENCE,  M.D. 


OTHERS    PREPARING. 

28 


The  Fiske  Fund  Prize  Essay  for  189O. 

THE 

SURGICAL  TREATMENT 

OF 

WOUNDS  AND  OBSTRUCTION 

OP    THE 

INTESTINES. 

BY 

EDWARD  MARTIN,  A.M.,  M.D., 

Clinical  Professor  of  Genito-Urinary  Diseases,  Instructor  in  Operative  Sur- 
gery, and  Lecturer  on  Minor  Surgery,  University  of  Pennsylvania; 
Surgeon  to  the  Howard  Hospital ;  Assistant  Surgeon  to  the 
University  Hospital,  etc.  etc. 
AND 

HOBART   A.  HARE,  M.D., 

Professor  of  Therapeutics,  Jefferson  Medical  College ;  Attending  Physician 
to  St.  Agnes'  Hospital. 


ILLUSTRATED. 


Price,  Cloth $2.00,  Net. 


"  In  presenting  this  Essay  upon  the  Surgical  Treatment  of  Wounds 
and  Obstruction  of  the  Intestines  to  the  Trustees  of  the  Fiske  Fund, 
it  is  proper  to  outline  the  scope  of  our  work,  and  to  state  "briefly  the 
facts  and  lines  of  original  research  upon  which  our  conclusions  are 
based.  For  over  two  years  we  have  made  experiments  in  the  labo- 
ratory upon  these  subjects,  and  have  carried  out  in  every  detail  all 
the  methods  and  modifications  of  operations  that  have  been  published 
or  which  have  occurred  to  us  in  the  course  of  our  own  studies.  .  .  . 
In  addition  to  the  original  work  involved  in  studying  so  important 
a  branch  of  surgery  as  the  one  before  us  (and  which  will  be  found 
represented,  graphically,  in  part  at  least  by  a  number  of  tracings), 
we  have  collected  and  placed  before  the  reader  what  we  believe  to  be 
the  fullest  statistics  yet  collected  upon  gunshot  wounds  of  the  abdo- 
men."— EXTRACT  FROM  PREFACE. 

29 


INDEX. 


PAGE 

ANNOUNCEMENT  .  .  . i 

AMERICAN  TEXT-BOOK  OF  SURGERY  .  .  .  .  2,  3 
VlERORDT  AND  STUART'S  MEDICAL  DIAGNOSIS  .  .  4 
KEATING'S  NEW  UNABRIDGED  DICTIONARY  or  MEDICINE  5 
SAUNDERS'  POCKET  MEDICAL  LEXICON  ....  6 
NANCREDE'S  ANATOMY  AND  MANUAL  OF  DISSECTION  .  7 
DESCHWEINITZ'S  DISEASES  OF  THE  EYE  ....  8 

GARRIGUE'S  DISEASES  OF  WOMEN 9 

NORRIS'  SYLLABUS  OF  OBSTETRICAL  LECTURES  .  .  9 
SAUNDERS'  POCKET  MEDICAL  FORMULARY  .  .  .10 
SAUNDERS*  SERIES  OF  QUESTION  COMPENDS  .  .  .11 

HARE'S  PHYSIOLOGY 12 

MARTIN'S  SURGERY  ....:...  13 

NANCREDE'S  ANATOMY 14 

WOLFF'S  CHEMISTRY 15 

ASHTON'S  OBSTETRICS 16 

SEMPLE'S  PATHOLOGY,  ETC 17 

MORRIS'  MATERIA  MEDICA 18 

MORRIS'  PRACTICE  OF  MEDICINE 19 

CRAGIN'S  GYNAECOLOGY 20 

STELWAGEN'S  DISEASES  OF  THE  SKIN  .  .  .  .21 

MARTIN'S  MINOR  SURGERY,  ETC 22 

SEMPLE'S  LEGAL  MEDICINE.  ETC 23 

JACKSON  AND  GLEASON'S  DISEASES  OF  EYE,  XOEI:,  AND 

THROAT -  24 

POWELL'S  DISEASES  OF  CHILDREN  ...  .25 

WOLFF'S  EXAMINATION  OF  URINE  ...  .26 

SAYRE'S  PRACTICE  OF  PHARMACY  .  .  27 

WORKS  IN  PREPARATION  AND  IN  PIM>-  .  .  .28 
MARTIN  AND  HARE'S  WOUNDS  AND  OBSTRUCTION  OF 

THE  INTESTINES -    29 

30 


THE 

CLIMATOLOGIST. 

A  MONTHLY  JOURNAL  OF  MEDICINE 


DEVOTED  TO  THE 


Relation  of  Climate,   Mineral  Springs,   Diet,  Pre- 
ventive Medicine,  Race,  Occupation,  Life 
Insurance  and  Sanitary  Science 
to  Disease. 


EDITED  BY 

JOHN    M.    KEATING,    M.  D. 
FREDERICK  A.  PACKARD,  M.  D.    CHAS.  F.  GARDINER,  M.  D. 


ASSOCIATE     EDITORS: 


NORMAN  BRIDGE,  M.D., 

Los  Angeles,  Cal. 
VINCENT  Y.  BOWD1TCH,  M.I). 

Boston,  Mass. 
SAML.  R.  BURROUGHS,  M.D., 

Raymond,  Tex. 
J.  WELLINGTON  BYEKS,  M.D., 

Charlotte,  N.  C. 
J.  M.  DACOSTA,  M.D., 

Philadelphia,  Pa. 
CHARLES  DENISON,  M.D., 

Denver,  Colo. 
GEORGE  DOCK,  M.[>., 

Galveston,  Texas. 
WM.  A.  EDWARDS,  M.D., 

San  Diego,  Cal. 
J.  T.  ESKRIDGE,  M.D., 

Denver,  Colo. 
S4MUEL  A.  FISK,  M.D.. 

Denver,  Colo. 
W.  H.  GEDDINGS,  M.D., 

Aiken,S.  C. 
JOHN  B.  HAMILTON,  M.D., 

Chicago,  111. 
T.  S.  HOPKINS,  M.D., 

Thomasviile,  Ga. 
FREDERICK  I.  KNIGHT.,  M.D., 

Boston,  Mass. 
R.  L.  MACDONNELL,  M.D., 

Montreal,  Canada. 


FRANCIS  MINOT,  M.D., Moaton.Muss. 
ALFRED  L.  LOOM1S,  M.I)  ,      , 

New  York  City. 
HENRY  M.  LYMAN,  M.D., 

Chicago,  Ills. 
WILLIAM  OSLER,  M.D., 

Baltimore,  Md. 
WILLIAM  PEPPER,  M.D., 

Philadelphia,  Pa. 
BOARDMAN  REED,  M.D., 

Atlantic  City.  N.  J. 
J.  REED,  JR.,  M.D., 

Colorado  Springs,  Colo. 
GEORGE  II.  ROHE,  M.D., 

Baltimore.  Md. 

KARL  VON  RUCK,  M.D., 

Asheville,  N.  C. 
FREDK.  C.  SHATTUCK,  M.I)., 

Boston,  Mass. 
S.  E.  SOLLY,  M.D., 

Colorado  Springs,  Colo. 
G.  B.  THORNTON,  M.D., 

Memphis,  Tenn. 
E.  L.  TRUDEAU,  M.D., 

Saranac  Luke,  N.  Y. 
J,  B.  WALKER,  M.   D., 

Philadelphia.  Pa. 

J.  P.  WALL,  M.D.,  Tampa,  Florida. 
,  M.D., 


JAMES  C.  WILSON, 


Philadelphia,  Pa. 


Yearly  Subscription  $2.OO.        Single  Numbers  2O  Cts. 

W.    B.    SAUNDERS,    Publisher, 

913  Walnut  Street,  Philadelphia,  Pa. 


EXTRACT    FROM    THE    INTRODUCTION    IN  THE    OPENING 

NUMBER  OF 

"THE  CLIMATOLOGIST." 

AUGUST,  1891. 


"  THE  object  of  this  JOURNAL  is  to  promote  original  investi- 
gation, to  publish  papers  containing  the  observations  and  ex- 
perience of  physicians  in  this  country  and  Europe  on  all  matters 
relating  to  CLIMATOLOGY,  MINERAL  SPRINGS,  DIET,  PREVENTIVE 
MEDICINE,  RACE,  OCCUPATION,  LIFE  INSURANCE,  AND  SANITARY 
SCIENCE — and  in  that  way  to  supply  the  means  by  which  the 
general  practitioner  and  the  public  at  large  will  become  better 
acquainted  with  the  diseases  of  this  country  and  Europe,  and 
better  armed  to  meet  the  requirements  of  their  prevention  or 
cure.  The  study  of  these  subjects  in  this  country  is  exciting 
great  and  increasing  interest,  and  all  admit  that,  from  the  little 
knowledge  already  possessed  of  its  resources,  possibly  every 
known  combination  of  atmospheric  condition,  soil,  altitude,  cli- 
mate, or  mineral  springs,  is  to  be  found  on  this  continent.  It  is 
confidently  expected  that  such  a.  journal \v\\\  receive  encourage- 
ment and  be  an  authority  upon  all  questions  which  are  included 
in  its  title. 

"  Original  papers  upon  diseases  of  localities — those  incident 
to  occupation,  race,  or  climate,  the  study  of  epidemics,  the 
questions  of  proper  food,  of  the  water  supply,  its  potability 
and  distribution,  matters  relating  to  drainage  and  diseases  de- 
pendent on  it — as  well  as  experimental  studies,  or  laboratory 
investigations  on  bacteriology,  will  form  a  prominent  portion 
of  the  material  presented  during  the  year,  and  it  is  to  be  hoped 
that  physicians  of  all  sections  of  the  country  will  send  papers 
upon  these  or  any  other  subjects  which  will  be  of  general  in- 
terest. 

"  Special  attention  will  also  be  paid  to  the  subject  of  health 
resorts,  descriptions  of  Sanitariums  with  special  reference  to 
their  suitability  to  certain  cases,  and  the  proper  selection  ot 
patients  likely  to  be  benefitted  by  them.  The  utmost  care  will 
be  taken  that  this  JOURNAL  shall  assume  and  maintain  the 
highest  scientific  character.  It  will  be  absolutely  independent 
in  its  principles— -fair  towards  all.  It  will  depend  for  its  main- 
tenance upon  the  support  given  to  it  by  the  prcfession,  as  it  is 
not  published  in  the  interest  of  any  special  section  or  clique." 


POCKET  MEDICAL  LEXICON; 


OR, 


Dictionary  of  Terms  and  Words  used  in  Medicine  and  Surgery, 

BY  JOHN  M.  KEATING,  M.V., 

Editor  of  "Cyclopedia  of  Diseases  of  Children,"  etc.;  Author  of  the 
"New  Pronouncing  Dictionary  of  Medicine," 


HENRY  HAMILTON, 

Author  of  "A  New  Translation  of  Virgil's  yEneid  into  English  Verse;' 
Co-author  of  a  "New  Pronouncing  Dictionary  of  Medicine." 


Price,  75  Cents,  Cloth.    $1.00,  Leather  Tucks. 


This  new  and  comprehensive 
work  of  reference  is  the  outcome 
of  a  demand  for  a  more  modern 
handbook  of  its  class  than  those 
at  present  on  the  market, which, 
dating  as  they  do  from  1855  to 
1884,  are  of  but  trifling  use  to 
the  student  by  their  not  con- 
taining the  hundreds  of  new 
words  now  used  in  current  lit- 
erature, especially  those  relat- 
ing to  Electricity  and  Bacteri- 
ology. 


y/b*_l 

q 

_2//?tf    _5<7* 

$0    — 

__/^     -.72 

do  — 

_/7tf      .« 

70    — 

-K*      -56 

60  — 

—  /fO     —48 

50  _ 

^I2Z      —40 

46   — 

-/04     _3l 

30    _ 

—  86      _  *f 

/'~ 

__  tf*      —^  /tf 

^0°    - 

_J/«     _/?° 

*•  /0    _,_ 

_/-/      -tf- 

—  2»"  — 

_J  -^       —  liT- 

o 


(From  Appendix  to  Medical  Lexicon.) 


Annals  of  (rt/nwcoloyy,  Phila- 
delpliia,  December,  189O. 

Saunders'  Pocket  Medical  Lexi- 
con—a very  complete  little  work, 
invaluable  to  every  student  of 
medicine.  It  not  only  contains  a 
very  large  number  of  words,  but 
also  tables  of  etymological  factors 
common  in  medical  terminology  ; 
abbreviations  used  in  medicine, 
poisons  and  antidotes,  etc. 


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MEDICAL  DIAGNOSIS. 


DR.  OSWALD    VIERORDT. 

Professor  of  Medicine  at  the  University  of  Heidelberg  ^formerly  Privat 
at  University  of  Leipzig  ;  Professor  of  Medicine  and  Director  of  the 
Medical  Polyclinic.at  the  University  of  Jena. 

Translated,  with  Additions,  from  the  Second  Enlarged  Germar 
Edition,  with  the  Author's  Permission. 

BY     : 

FRANCIS  H.  STUART,  A.M.,  M.D., 

Member  of  the  Medical  Society  of  the  County  of  Kings,  N.  Y.  ;  Fellow  of  the 

New  York  Academy  of  Medicine  ;   Member  of  the  British 

Medical  Association,  etc. 


NUMEROUS  COLORED  AND  WOOD  ENGRAVINGS. 
Price,  Cloth,  $4.00;  Sheep,  $5.OO. 


In  this  work,  as  in  no  other  hitherto  published,  are  given  fall  and 
accurate  explanations  of  the  phenomena  observed  at  the  beds! 
is  distinctly  a  Clinical  work  by  a  master  teacher,  characterized  by  thor- 
oughness, fulness,  and  accuracy. 

It  is  a  mine  of  information  upon  the  points  that  are  so 
often  passed  over  without  explanation. 

The  student  who  is  familiar  with  its  contents  will  have  a  sound  foun- 
dation for.  the  practice  of  his  profession. 

The  author  gives  a  complete  though  brief  presentation  of  the  Micro- 
organisms,  whose  recognition  and  discrimination  are  made  pos^ 
cultivation,  and  inoculation,  and  which,  through  the  labors  of  those 
eminent  bacteriologists,    PASTEUR,   KOCH,   and   others,  h, 
made  such  marked' changes  in  the  application  of  IVIM-M! 
the  cure  of  disease. 


